Norgen Gel Extraction Kit. Norgen PCR Purification Kit. Norgen Plasmid MiniPrep Kit

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2 Using innovative nucleic acid resin technology, Norgen s kits provide fast processing time and high recovery for sample clean-up and preparation - all in a convenient spin column format. Choose from five different kits, available in a variety of sizes and formats. Norgen Gel Extraction Kit The Norgen DNA Gel Extraction Kit enables rapid purification of fractionated DNA fragments in agarose gels. Norgen PCR Purification Kit The Norgen PCR Purification Kit enables rapid purification of amplified DNA products from PCR mixes. Norgen Plasmid MiniPrep Kit The Norgen Plasmid MiniPrep Kit enables rapid analytical scale preparation of high quality plasmid DNA.

3 Norgen Plasmid MaxiPrep Kit The Norgen Plasmid MaxiPrep Kit provides a rapid method for the preparation of large quantities of high quality plasmid DNA. Norgen Total RNA Purification Kit Norgen s Total RNA Purification Kit provides a rapid method for the isolation and purification of total RNA from cultured animal cells, bacterial cells, blood and small tissue samples. In addition to DNA sample preparation kits, Norgen also offers DNA and RNA molecular weight markers for use in electrophoresis. DNA Ladders and RNA Ladders Norgen s DNA Ladders are ideal for accurate size determination and quantification of DNA using ethidium bromide stained agarose gels. Norgen s RNA Ladders are ideal for precise sizing of RNA molecules using native or denaturing gels.

4 Norgen DNA Gel Extraction Kit The Norgen DNA Gel Extraction Kit enables rapid purification of fractionated DNA fragments in agarose gels. Purification is based on spin column chromatography using Norgen s proprietary resin as the separation matrix. Chaotropic salts are used to melt agarose gel slices and also allow reversible binding of the target DNA to the matrix. The kit effectively removes gel contaminants, allows for buffer exchange and yields high quality DNA appropriate for many downstream applications. The Norgen DNA Gel Extraction Kit is able to recover DNA ranging in size from 100 bp to 10,000 bp. Each column has a binding capacity of 10 µg, and the typical purification yield is up to 90% of the input amount. Norgen DNA Gel Extraction Kit Benefits Fast and easy processing Large column reservoir High recovery Recovered DNA is suitable for downstream applications Useable with a variety of agarose percentages Long kit shelf life Rapid spin-column format allows for the processing of multiple samples in 30 minutes. 1mL column reservoir capacity allows for the processing of large gel slices in a single spin. Recovery of DNA fragments up to 90% for inputs of > 1 µg. Purified DNA is fully compatible with restriction digestions, ligations, and other downstream applications. Kit is compatible with agarose gel concentrations of up to 4%. Kit has a shelf life of 1 year.

5 Yield (nanogram) N Y Z M C1 N1 Y1 C2 N2 Y2 C3 N3 Y3 Figure 1. Efficient Recovery from Agarose Gels A 500bp fragment was purified from a 1% agarose gel using the Norgen DNA Gel Extraction Kit (N) and two other commercial kits (Y and Z). The Norgen kit shows a significantly higher efficiency compared to the competitor kits. Figure 2. Broad Size Range of Target DNA A 261bp fragment (Lane C1), a 630bp fragment (Lane C2) and a 1,058bp fragment (Lane C3) were recovered using both Norgen s kit (N1-N3) and a competitor s kit (Y1-Y3). M Input A B C Figure 3. Full Compatibility with Restriction Digests In order to demonstrate the purity of gel extracted DNA fragments, the Norgen DNA Gel Extraction Kit was used to purify a 1,200bp PCR product, which was subsequently digested with BamHI (Lane A), HindIII (Lane B) and PvuII (Lane C). Norgen DNA Gel Extraction Kit Contents 1. Binding Solution 2. Wash Solution 3. Elution Buffer 4. Micro Spin Columns 5. Elution Tubes 6. Short Protocol Card 7. User Manual Customer-Supplied Reagents and Equipment Benchtop microcentrifuge Micropipettors 95% ethanol Isopropanol Cat # Description Quantity Norgen DNA Gel Extraction Kit 50 samples Norgen DNA Gel Extraction Kit 100 samples

6 Norgen PCR Purification Kit The Norgen PCR Purification Kit enables rapid purification of amplified DNA products from PCR mixes. Purification is based on spin-column chromatography using Norgen s proprietary resin as the separation matrix. Norgen s resin binds DNA under high salt concentrations and releases the bound DNA under low salt and slightly alkali conditions. This kit is able to effectively remove PCR by-products including primers, dimers, enzymes, unincorporated nucleotides and mineral oil from the desired PCR product. The Norgen PCR Purification Kit is able to purify amplified DNA ranging in size from 100 bp to 15,000 bp. Each column has a binding capacity of 10 µg, and the typical purification yield is up to 95% of the input amount. Norgen PCR Purification Kit Benefits Fast and easy processing Purification from all PCR by-products High recovery Recovered DNA is suitable for downstream applications Can be used as an alternative to organic extraction and ethanol precipitation Long kit shelf life Rapid spin-column format allows for the processing of multiple samples in 15 minutes. The kit removes primer-dimers, primers, mineral oil, unincorporated nucleotides, enzymes and salts. Recovery of PCR products is up to 95% of input amount. Purified DNA is fully compatible with restriction digestions, ligations, labeling and sequencing. The kit can also be used to clean-up various enzymatic reactions, including digestions and Klenow reactions. Kit has a shelf life of 1 year.

7 Yield (nanogram) N1 N2 N3 Y1 Y2 Y3 M U P U P U P U P U P U P Figure 1. High Yield Purification The high yield of the Norgen PCR Purification Kit is illustrated by purifying a 500bp PCR-amplified product in a reaction mixture and comparing the recovery with a competitor. The results are shown in Figure 1, which depicts the yields of triplicates (N1 to N3 for Norgen, and Y1 to Y3 for the competitor). Figure 2. Removal of Primer-Dimers Primer-dimers present in PCR reactions are effectively removed during the purification process with the Norgen PCR Purification Kit. An additional wash with guanidine hydrochloride is added in the procedure to remove greater than 85% of primer dimers. The figure shows alternating lanes containing unprocessed (U) and processed (P) PCR reaction products. Norgen PCR Purification Kit Contents 1. Binding Solution 2. Wash Solution 3. Elution Buffer 4. Micro Spin Columns 5. Elution Tubes 6. Short Protocol Card 7. User Manual Customer-Supplied Reagents and Equipment Benchtop microcentrifuge Micropipettors 95% ethanol 4M guanidine hydrochloride (optional) Cat # Description Quantity Norgen PCR Purification Kit 50 samples Norgen PCR Purification Kit 100 samples

8 Norgen Plasmid MiniPrep Kit The Norgen Plasmid MiniPrep Kit enables rapid analytical scale preparation of high quality plasmid DNA. Purification is based on spin column chromatography using Norgen s proprietary resin as the separation matrix. Norgen s resin binds DNA under high salt concentrations and releases the bound DNA under low salt and slightly alkali conditions. The kit purifies DNA without alcohols, a significant improvement over other techniques to achieve full compatibility with alcoholsensitive applications. The purified DNA is free of genomic DNA, RNA and cellular proteins and is suitable for many downstream applications. The Norgen Plasmid MiniPrep Kit is able to purify plasmids up to 13,000 bp. Each column has a binding capacity of 25 µg, and the typical purification yield is up to 19 µg from 1.5 ml of bacterial culture. Norgen Plasmid MiniPrep Kit Benefits Fast and easy processing High binding capacity of columns High recovery Recovered plasmid DNA is suitable for downstream applications Alcohol-free protocol Long kit shelf life Rapid spin-column format allows for the processing of multiple samples in 30 minutes. The binding capacity of the columns in the Plasmid MiniPrep Kit is 25 µg. Up to 19 µg of plasmid DNA can be purified from 1.5 ml of bacterial culture. Purified plasmid DNA is fully compatible with restriction enzyme digestion and sequencing. No alcohols are used during the plasmid purification process, thus the DNA is compatible with alcohol sensitive downstream applications. Kit has a shelf life of 1 year.

9 Yield (microgram) N1 N2 N3 Y1 Y2 Y3 Z1 Z2 Z3 Figure 1. High DNA Yield from Small Cultures Plasmid DNA from bacterial cultures grown overnight was prepared with the Norgen Plasmid Mini- Prep Kit and compared with two other commercial kits. The yield for each of the triplicates is shown on the graph (N1 to N3 for Norgen, and Y1 to Y3 and Z1 to Z3 for competitor kits). Figure 2. High-Accuracy Sequencing of DNA Product Sequencing of purified plasmid DNA as a template resulted in an accuracy of % over a 950 bp contiguous sequence. 15 minute digest 30 minute digest 60 minute digest MR N Y Z C N Y Z C N Y Z C MR Figure 3. Full Compatibility with Restriction Digests Purified plasmid DNA is highly digestible with restriction enzymes, usually requiring less time to digest completely than DNA purified with other kits as shown in time-course digestion experiments. In Figure 3 digestions were performed with HindIII, and N = Norgen DNA, Y = competitor 1, Z = competitor 2 and C = control DNA. Norgen Plasmid MiniPrep Kit Contents 1. Resuspension Buffer 2. Lysis Solution 3. Binding Solution 4. RNAse A 5. Wash Solution 6. Elution Buffer 7. Micro Spin Columns 8. Elution Tubes 9. Short Protocol Card 10. User Manual Customer-Supplied Reagents and Equipment Benchtop microcentrifuge Micropipettors 1.5 ml microcentrifuge tubes Cat # Description Quantity Norgen Plasmid MiniPrep Kit 50 samples Norgen Plasmid MiniPrep Kit 100 samples

10 Norgen Plasmid MaxiPrep Kit The Norgen Plasmid MaxiPrep Kit provides a rapid method for the preparation of large quantities of high quality plasmid DNA. Purification is based on spin column chromatography using Norgen s proprietary resin as the separation matrix. Norgen s resin binds DNA under high salt concentrations and releases the bound DNA under low salt and slightly alkali conditions. The kit purifies DNA without alcohols, a significant improvement over other techniques to achieve full compatibility with alcohol-sensitive applications. The purified DNA is free of genomic DNA, RNA and cellular proteins and is suitable for many downstream applications. The Norgen Plasmid MaxiPrep Kit is able to purify up to 1.5 mg of plasmid DNA from 150 ml of bacterial culture containing a high copy number plasmid. Norgen Plasmid MaxiPrep Kit Benefits Fast and easy processing High binding capacity of columns High recovery Recovered plasmid DNA is suitable for downstream applications Alcohol-free protocol Long kit shelf life Rapid spin-column format allows for the processing of multiple samples in 2 hours. The binding capacity of the columns in the Plasmid MaxiPrep Kit is 1.5 mg. Up to 1.5 mg of plasmid DNA can be purified from 150 ml of bacterial culture. Purified plasmid DNA is fully compatible with restriction enzyme digestions, sequencing and transfections. No alcohols are used during the plasmid purification process, thus the DNA is compatible with alcohol sensitive downstream applications. Kit has a shelf life of 1 year.

11 Yield (micrograms) N1 N2 N3 Y1 Y2 Y3 Z1 Z2 Z3 M U A B C Figure 1. High DNA Yield from 150 ml Cultures Plasmid DNA from bacterial cultures grown overnight was prepared with the Norgen Plasmid MaxiPrep Kit and compared with two other commercial kits. The yield for each of the triplicates is shown on the graph (N1 to N3 for Norgen, and Y1 to Y3 and Z1 to Z3 for competitor kits). Figure 2. Full Compatibility with Restriction Digests DNA isolated with the Norgen Plasmid MaxiPrep kit is easily digestible, often requiring less than 1 hour for full digestion. In Figure 2, a 9,309 bp plasmid was purified and digested with BamHI (Lane A), HindIII (Lane B), and SmaI (Lane c). Lane U is uncut plasmid, and Lane M is the Norgen Ultra- Ranger 1kb DNA ladder. Norgen Plasmid MaxiPrep Kit Contents 1. Resuspension Buffer 2. Lysis Solution 3. Neutralization Solution 4. Binding Solution 5. RNAse A 6. RNAse T1 7. Wash Solution 8. Elution Buffer 9. Maxi Spin Columns 10. Filter Columns 11. Elution Tubes 12. Short Protocol Card 13. User Manual Customer-Supplied Reagents and Equipment Centrifuge with a swinging bucket rotor capable of 3000 x g Centrifuge capable of x g Micropipettors Cat # Description Quantity Norgen Plasmid MaxiPrep Kit 4 samples

12 Total RNA Purification Kit Norgen s Total RNA Purification Kit provides a rapid method for the isolation and purification of total RNA from cultured animal cells, bacterial cells, blood and small tissue samples. Purification is based on spin column chromatography using Norgen s proprietary resin as the separation matrix. Norgen s resin binds RNA in a manner that depends on ionic concentrations. The RNA is preferentially purified from other cellular components such as genomic DNA or proteins. The purified RNA is of the highest integrity, and can be used in a number of downstream applications. Each spin column in the Norgen Total RNA Purification Kit has a binding capacity of up to 15 µg of RNA. RNA can be isolated from 1 to 10 6 cells growing in cell culture, from bacterial cells and blood, and from up to 2 mg of tissue. It should be noted that total RNA can be isolated from larger amounts of tissue, however the sample must be filtered first to prevent clogging of the column. The purified RNA can be used for reverse transcription PCR, real time PCR, Northern blotting, RNase protection, primer extension and expression array assays. Norgen Total RNA Purification Kit Benefits Fast and easy processing Rapid spin-column format allows for the processing of 10 samples in 25 minutes. High binding capacity of columns The binding capacity of the columns in the Total RNA Purification Kit is up to 15 µg. Isolate total RNA from a variety of sources RNA can be isolated from a variety of cell sources, including animal cells growing in a monolayer, suspension or lifted cells, bacterial cells, blood and small tissue samples. (Figure 3). Isolate total RNA from very small samples Recovered RNA is suitable for downstream applications Long kit shelf life Total RNA has been isolated and detected from as little as a single cell (Figure 1). Purified RNA can be used in a number of downstream applications including real-time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array analysis require the use of intact RNA. Kit has a shelf life of 1 year.

13 M A B C D M Figure 1. Isolation and detection of RNA from as little as a single animal cell. RT-PCR was used to detect RNA isolated from 293 HEK cells using the Norgen Total RNA Purification Kit. M is the marker lane, while 10 6 to 1 corresponds to the number of cells lysed in order to isolate total RNA. Figure 3. Total RNA isolated from various sources was analyzed on an RNA Labchip using the Agilent 2100 BioAnalyzer. In all cases, 1 µl of total RNA was analyzed. Lane M is the marker, while Lanes A-D are total RNA isolated from 20 µl of blood (Lane A), 12 mg of kidney (Lane B), 8 mg of lung (Lane C), and 1 mg of spleen (Lane D). The high quality of the isolated RNA is demonstrated by the presence of 2 clear bands, representing the 18S and 28S ribosomal RNAs. Figure 2. RNA isolated using the Norgen Total RNA Purification Kit is of the highest integrity and quality. One µl of total RNA isolated from cultured 293 HEK cells was analyzed on an RNA Labchip using the Agilent 2100 BioAnalyzer. Clear peaks can be observed for both the 18S and 28S ribosomal RNAs. Norgen Total RNA Purification Kit Contents 1. Lysis Solution 2. Activation Solution 3. Wash Solution 4. Elution Buffer 5. Micro Spin Columns 6. Elution Tubes 7. Short Protocol Card 8. User Manual Customer-Supplied Reagents and Equipment Benchtop microcentrifuge Micropipettors Cat # Description Quantity Norgen Total RNA Purification Micro Kit 50 samples

14 DNA Ladders Norgen s DNA Ladders are ideal for accurate size determination and quantification using agarose gels stained with ethidium bromide. Features: Quantitative Stable at room temperature Ready-to-use Higher intensity reference bands Discrete, well-spaced bands Full range of markers to suit all PCR, cloning and research needs Cat # Description Quantity MiniSizer 50bp DNA Ladder 100 loads PCRRanger 100bp DNA Ladder 100 loads PCRSizer 100bp DNA Ladder 100 loads FastRunner DNA Ladder 100 loads LowRanger 100bp DNA Ladder 100 loads Cat # Description Quantity CloneSizer 100bp DNA Ladder 100 loads FullRanger 100bp DNA Ladder 100 loads HighRanger Plus 100bp DNA Ladder 100 loads HighRanger 1kb DNA Ladder 100 loads UltraRanger 1kb DNA Ladder 100 loads RNA Ladders Norgen s RNA Ladders are ideal for precise sizing of RNA molecules using native or denaturing gels. Features: Easy to reconstitute No need for staining or destaining Stable Higher intensity reference bands Cat # Description Quantity b RNA Ladder, Lyophilized 50 loads kb RNA Ladder, Lyophilized 50 loads

15 Corporate Headquarters 344 Merritt St. St. Catharines, Ontario CANADA L2T 1K6 Phone (905) or Toll Free in North America: Fax (905) Ordering Information Norgen Biotek offers several ways to place orders 24 hours a day. Phone: (905) or Toll Free in North America: Website: orders@norgenbiotek.com Fax: (905) To locate a distributor in your area, please contact Norgen Biotek. Technical Support Contact our Technical Support Team between the hours of 8:30 and 5:30 (Eastern Standard Time) at (905) or Toll Free at Technical support can also be obtained from our website ( or through at techsupport@norgenbiotek.com. Trademarks and Disclaimers Norgen s nucleic acid purification technology is covered by U.S. Patents 6,177,278, 6,291,248, and the pending U.S. Patent 10/379,383. Norgen s nucleic acid purification technology is also covered by a number of foreign patents including U.K. Patent GB , Australian Patent , the pending Japanese patent ,378, and pending German patent Some product compositions are proprietary and undisclosed. The following trademarks also used in this publication are the property of the companies listed below. LabChip is a registered trademark of Caliper Technologies Corp. Agilent 2100 Bioanalyzer is a registered trademark of Agilent Technologies, Inc.

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18 Using innovative protein resin technology, ProteoSpin kits provide fast processing time and high protein recovery for sample clean-up and preparation - all in a convenient spin column format. Choose from seven ProteoSpin kits, available in a variety of sizes and formats. CBED (Concentration, Buffer Exchange and Desalting) Micro Kit CBED (Concentration, Buffer Exchange and Desalting) Maxi Kit Fast and simple procedure for concentration, buffer exchange and desalting of small and large protein samples. Detergent Clean-up Micro Kit Detergent Clean-up Maxi Kit Effective removal of SDS, Triton X-100 and other detergents from small and large protein samples. Inclusion Body Isolation Micro Kit Facilitates the screening of E. coli clones that express recombinant proteins in inclusion bodies.

19 Urine Protein Concentration Micro Kit Fast and simple procedure for concentrating dilute solutions of urine proteins from small volumes of urine. Serum Depletion Kit Effective depletion of abundant serum proteins including albumin, α-antitrypsin, transferin and haptoglobin from serum and plasma samples. In addition to Proteospin protein sample preparation kits, Norgen also offers protein ladders for use in electrophoresis. ProteoLadder Protein Ladders The ProteoLadders are ideal for accurate molecular weight sizing of proteins by SDS- PAGE gels stained with Coomassie blue.

20 ProteoSpin CBED (Concentration, Buffer Exchange and Desalting) Micro Kit The ProteoSpin CBED Micro Kit provides a fast and simple procedure for concentration, buffer exchange and desalting of protein samples. The kit is based on spin-column chromatography using Norgen s proprietary protein resin as an ion exchanger. Salts such as MgCl 2, NaCl, KCl, CaCl2, LiCl, and CsCl have poor affinities for Norgen s resin and are easily removed from the sample. The elution is performed with choices of several different buffer solutions. Each spin column can process 2-50 µg of protein from up to 5 ml of solution (Table 1). The kit contains complete solutions for the preparation of both acidic and basic proteins. The ProteoSpin CBED Micro Kit provides a convenient method for preparing proteins before running downstream proteomic applications including mass spectrometry, SDS-PAGE, isoelectric focusing, x-ray crystallography and NMR spectroscopy. ProteoSpin CBED Micro Kit Benefits Fast and easy processing No mixing or formulation No molecular weight cutoff Proteins bind while salts are discarded in flowthrough Long kit shelf life Suitable for downstream applications Efficiently process up to 12 samples in only 20 minutes using an easyto-use protocol. Rapid spin-column chromatography format with yields up to 99% of input amount. Solutions and protocols are provided for both acidic and basic proteins with no need for formulation. Based on an ion exchange mechanism, a broad size range of proteins from peptides to antibodies can be processed. Norgen s resin has an inherent low affinity for salt but high affinity for proteins, providing salt removal and buffer exchange simultaneously (Table 2). 1 year Final elution performed with volatile or non-volatile buffers suitable for: Mass spectrometry (Figure 1) SDS-PAGE Isoelectric focusing X-ray crystallography NMR spectroscopy

21 A B Figure 1. Samples from a biopsy lysate were initially concentrated using a C18 ZipTip and analyzed using MALDI-TOF (Panel A). Addition of a gel filtration step followed by concentration using the ProteoSpin CBED Micro Kit allowed improved signal detection (Panel B). Data courtesy of Eric Yang (Ph.D.) and Michael Siu (Ph.D.), York University, Toronto. Table 1. Examples of BSA and IgG Concentration Using the CBED Micro Kit Volume Concentration 2,000 µl to 50 µl (40X) Mass Concentration (µg/µl) 50 µg/2,000 µl to µg/50 µl µg/µl to µg/µl (32X-40X) Table 2. Representative Recovery and Salt Reduction Using the ProteoSpin CBED Micro Kit Salt (300 ppm) % BSA Recovery % Salt Reduction MgCl NaCl KCl CaCl LiCl CBED Micro Kit Contents Each CBED Micro Kit provides essential components for use with the provided protocols. 1. Column Activation and Wash Buffer for Acidic Proteins 2. ph Binding Buffer for Acidic Proteins 3. Column Activation and Wash Buffer for Basic Proteins 4. ph Binding Buffer for Basic Proteins 5. Elution Buffer 6. Neutralizer 7. Micro Spin Columns 8. Elution Tubes 9. Short Protocol Card 10. Application Manual Customer-Supplied Reagents and Equipment Benchtop microcentrifuge Micropipettors ph indicator paper Other elution buffers (optional) Cat # Description Quantity ProteoSpin CBED Micro Kit 25 samples ProteoSpin CBED Micro Kit 50 samples

22 ProteoSpin CBED (Concentration, Buffer Exchange and Desalting) Maxi Kit The ProteoSpin CBED Maxi Kit provides a fast and simple procedure for concentrating large volumes of dilute protein solutions for buffer exchange, and for removing different types of salts from protein samples. The kit is based on spincolumn chromatography using Norgen s proprietary protein resin as an ion exchanger. Salts such as MgCl 2, NaCl, KCl, CaCl2, LiCl, and CsCl have poor affinities for Norgen s resin and are easily removed from the sample. The elution is performed with choices of several different buffer solutions. Each spin column can process mg of protein from up to 100 ml of solution. The kit contains complete solutions for the preparation of both acidic and basic proteins. The ProteoSpin CBED Maxi Kit can be used to prepare protein samples for structural analysis when larger amounts of proteins are needed, such as x-ray crystallography, NMR spectroscopy and other applications. ProteoSpin CBED Maxi Kit Benefits Fast and easy processing No mixing or formulation No molecular weight cutoff Proteins bind while salts are discarded in flowthrough Long kit shelf life Suitable for downstream applications Efficient processing of multiple samples in about 30 minutes using an easy-to-use protocol. Solutions are provided for both acidic and basic proteins with no need for formulation. Based on an ion exchange mechanism, a broad size range of proteins (from peptides to large proteins) can be processed. Norgen s resin has an inherent low affinity for salt but high affinity for proteins, providing salt removal and buffer exchange simultaneously. 1 year Final elution performed with volatile or non-volatile buffers suitable for: Mass spectrometry SDS-PAGE Isoelectric focusing X-ray crystallography NMR spectroscopy

23 2-Elution Total (microgram) Input BSA (microgram) Figure 1. The columns used in the ProteoSpin CBED Maxi Kit have a high binding capacity. The saturation point of the columns is 8 mg of protein, as can be seen in the binding of increasing amounts of BSA. The maximum recovery efficiency for BSA was found to be 89% in the linear range. Table 1. Example of BSA Concentration Using the CBED Maxi Kit CBED Maxi Kit Contents Each CBED Maxi Kit provides essential components for use with the provided protocols. 1. Column Activation and Wash Buffer for Acidic Proteins 2. ph Binding Buffer for Acidic Proteins 3. Column Activation and Wash Buffer for Basic Proteins 4. ph Binding Buffer for Basic Proteins 5. Elution Buffer 6. Neutralizer 7. Maxi Spin Columns 8. Elution Tubes 9. Short Protocol Card 10. Application Manual Customer-Supplied Reagents and Equipment Benchtop centrifuge Micropipettors ph indicator paper Other elution buffers (optional) Volume Concentration 20 ml to 4 ml (5X) Mass Concentration (µg/µl) 8 mg/20 ml to 7.89 mg/4 ml 0.4 µg/µl to 1.97 µg/µl (5X) Cat # Description Quantity ProteoSpin CBED Maxi Kit 4 samples

24 ProteoSpin Detergent Clean-up Micro Kit Detergents are extensively used to prepare protein samples. However, detergents must often be removed prior to downstream analysis because of their undesirable effects. These include extraneous peaks in mass spectrometry, artifacts with chromatography and electrophoresis, interference with microinjection into cells and interference with protein immunization. The ProteoSpin Detergent Clean-up Micro Kit provides a fast and simple procedure for the effective removal of SDS, Triton X-100 and other detergents from small samples of protein. The kit employs spin-column chromatography using Norgen s proprietary protein resin as an ion exchanger. Each column is capable of processing 2 50 µg of either acidic or basic protein. ProteoSpin Detergent Clean-up Micro Kit Benefits Fast and easy processing In contrast to longer dialysis procedures, this easy-to-use procedure allows processing of up to 12 samples in only 20 minutes. Detergent removal and concentration is done simultaneously. No detergent carryover Useable with a variety of detergents Effective detergent removal Long kit shelf life High recovery Suitable for downstream applications Unlike HPLC procedures, each sample is processed individually using its own spin tube, resulting in no carryover or column bleed from sample to sample. The kit can be used to remove detergents including SDS, Triton X- 100, CHAPS, NP-40, and Tween 20. Removes more than 95% of detergents (Table 1) for many protein samples, allowing tryptic digestion (Figure 1A, B); contrasting samples using molecular weight cut-off membranes show residual detergent that can interfere with trypsin digestion (Figure 1C). 1 year Protein recoveries of 80 to 95% (using initial detergent concentrations of 0.1%, Figure 2) for many proteins. Final elution suitable for: Mass spectrometry SDS-PAGE Proteolytic digestion Isoelectric focusing X-ray crystallography NMR spectroscopy

25 Table 1. Efficient Removal of SDS Sample A600 % SDS Remaining % SDS Removal mg SDS Removed Control (water) % SDS ProteoSpin sample 1 ProteoSpin sample % 99.98% % 99.98% Figure 1. A) BSA sample digested with trypsin without removal of Triton X-100. Only Triton peaks observed. B) BSA sample processed using Proteo- Spin Detergent Clean-up Micro Kit, then digested, showing BSA tryptic proteins. C) BSA sample processed using 3,000 MW cutoff membrane prior to tryptic digestion still shows inhibition of trypsin digestion due to residual Triton X-100. Data courtesy of Eric Yang (Ph.D.) and Michael Siu (Ph.D.) York University, Toronto % 90.0% 80.0% 70.0% 60.0% 50.0% 40.0% 30.0% 20.0% 10.0 % 0.0% 0%SDS 0.1%SDS 0.3%SDS 0.6%SDS 1.0%SDS SDS Figure 2. Solutions containing 50 µg BSA and 0-1.0% SDS were processed using the ProteoSpin Detergent Clean-up Micro Kit to remove SDS. High protein recovery was obtained with original SDS concentrations as high as 1%. Detergent Clean-up Micro Kit Contents Each Detergent Clean-up Micro Kit provides essential components for use with the provided protocols. 1. Column Activation and Wash Buffer for Acidic Proteins 2. ph Binding Buffer for Acidic Proteins 3. Column Activation and Wash Buffer for Basic Proteins 4. ph Binding Buffer for Basic Proteins 5. Elution Buffer 6. Neutralizer 7. Micro Spin Columns 8. Elution Tubes 9. Short Protocol Card 10. Application Manual Customer-Supplied Reagents and Equipment Benchtop microcentrifuge Micropipettors ph indicator paper Other optional elution buffers Isopropanol Sterile, deionized water or Milli-Q water Cat # Description Quantity ProteoSpin Detergent Clean-up Micro Kit 25 samples ProteoSpin Detergent Clean-Up Micro Kit 50 samples

26 ProteoSpin Detergent Clean-up Maxi Kit Detergents are extensively used to prepare protein samples. However, detergents must often be removed prior to downstream analysis because of their undesirable effects. These include extraneous peaks in mass spectrometry, artifacts with chromatography and electrophoresis, interference with microinjection into cells and interference with protein immunization. The ProteoSpin Detergent Clean-up Maxi Kit provides a fast and simple procedure for the effective removal of SDS, Triton X-100 and other detergents from protein samples. The kit employs spin-column chromatography using Norgen s proprietary protein resin as an ion exchanger. Each column is capable of processing mg of either acidic or basic protein. ProteoSpin Detergent Clean-up Maxi Kit Benefits Fast and easy processing In contrast to longer dialysis procedures, this easy-to-use procedure allows processing of a number of samples in about 40 minutes. No detergent carryover Useable with a variety of detergents Effective detergent removal High recovery Long kit shelf life Suitable for downstream applications Unlike HPLC procedures, each sample is processed individually using its own spin tube, resulting in no carryover or column bleed from sample to sample. The kit can be used to remove detergents including SDS, Triton X- 100, CHAPS, NP-40, and Tween 20. Removes more than 95% of detergents for many protein samples, allowing tryptic digestion without residual detergents that interfere with trypsin digestion. Protein recoveries of up to 100% (using initial detergent concentrations of 0.1% and 1.0%, Table 2) for many proteins 1 year Final elution suitable for: Mass spectrometry SDS-PAGE Proteolytic digestion Isoelectric focusing X-ray crystallography NMR spectroscopy

27 Table 1. Efficient Removal of SDS Sample A600 % SDS Remaining Control (water) 0.1% SDS Sample 1 (0.1% SDS) Sample 2 (1.0% SDS) % SDS Removal mg SDS Removed % 99.99% % 99.99% Removal of SDS with the ProteoSpin Detergent Clean-up Maxi Kit is greater than 99%, with up to 50 mg of SDS being removed. Detergent Clean-up Maxi Kit Contents Each Detergent Clean-up Maxi Kit provides essential components for use with the provided protocols. 1. Column Activation and Wash Buffer for Acidic Proteins 2. ph Binding Buffer for Acidic Proteins 3. Column Activation and Wash Buffer for Basic Proteins 4. ph Binding Buffer for Basic Proteins 5. Elution Buffer 6. Neutralizer 7. Maxi Spin Columns 8. Elution Tubes 9. Short Protocol Card 10. Application Manual Table 2. High Recovery of Proteins from an SDS- Containing Solution Input Amount (mg) % SDS Eluted Amount (mg) % Recovery Customer-Supplied Reagents and Equipment 1 mg Lysosyme 0.1% 0.94 mg 94% 2 mg Lysozyme 1.0% 2 mg 100% 2 mg BSA 0.1% 2 mg 100% 4 mg BSA 1.0% 3.7 mg 92.5% The recovery of both acidic and basic proteins from solutions containing SDS is high using the ProteoSpin Detergent Clean-up Maxi Kit. Recoveries of up to 100% are often observed, as can be seen in Table 2. Benchtop centrifuge Micropipettors ph indicator paper Other optional elution buffers Isopropanol Sterile, deionized water or Milli-Q water Cat # Description Quantity ProteoSpin Detergent Clean-up Maxi Kit 4 samples

28 ProteoSpin Inclusion Body Isolation Micro Kit The ProteoSpin Inclusion Body Isolation Micro Kit facilitates the screening of E. coli clones that express recombinant proteins in inclusion bodies. The kit includes reagents specially formulated to achieve rapid and high-quality purification of inclusion bodies using three processes: 1. Lysis of bacterial cells to release inclusion bodies in solid form 2. Solubilization of purified inclusion bodies 3. Purification of the recombinant protein using SiC spin column chromatography Downstream applications include SDS-PAGE analysis, refolding experiments, mass spectrometry, further purification and scale up. The ProteoSpin Inclusion Body Isolation Micro Kit employs spin-column chromatography using Norgen s proprietary protein resin as an ion exchanger. Each spin column can isolate up to 50 µg of either acidic or basic protein from 1.5 ml of bacterial culture. ProteoSpin Inclusion Body Isolation Micro Kit Benefits Three kits in one for acidic and basic proteins Gentle disruption procedure provides high purity final product. The kit provides essential reagents for cell disruption, inclusion body solublization and purification using spin column chromatography. The kit includes solutions and protocols for use with either acidic or basic proteins (Figures 1 and 2). Cell lysis is accomplished through non-ionic detergent chemical disruption. The use of a needle and syringe reduces viscosity and facilitates purification, producing proteins that are often >95% pure. Proteins are then ready for SDS-PAGE, 2D gels, mass spectrometry analysis or other procedures. Convenient process volume Fast and easy processing Long kit shelf life Suitable for downstream applications Analyze 1.5 ml of microbial cell culture yielding 2-50 µg of protein. Process up to 12 samples in only 60 minutes. The kit uses an enzyme-free process with stable solutions for longer shelf life. Final elution performed with volatile or non-volatile buffers suitable for: Mass spectrometry SDS-PAGE Refolding experiments

29 Inclusion Body Isolation Micro Kit Contents Each Inclusion Body Isolation Micro Kit provides essential components for use with the provided protocols. Figure 1. Isolation of a recombinant 50 kda chimeric protein (acidic) using the ProteoSpin Inclusion Body Isolation Micro Kit. 1. Cell Lysis Reagent 2. Inclusion Body Solubilization Reagent 3. Column Activation and Wash Buffer for Acidic Proteins 4. ph Binding Buffer for Acidic Proteins 5. Column Activation and Wash Buffer for Basic Proteins 6. ph Binding Buffer for Basic Proteins 7. Elution Buffer 8. Neutralizer 9. Micro Spin Columns 10. Collection Tubes 11. Elution Tubes 12. Syringes 13. Needles 14. Short Protocol Card 15. Application Manual Figure 2. Isolation of recombinant RNaseA (basic protein) using the ProteoSpin Inclusion Body Isolation Micro Kit. Customer-Supplied Reagents and Equipment Benchtop microcentrifuge Micropipettors Sterile, deionized water or Milli-Q water Cat # Description Quantity ProteoSpin Inclusion Body Isolation Micro Kit 20 samples ProteoSpin Inclusion Body Isolation Micro Kit 50 samples

30 ProteoSpin Urine Protein Concentration Micro Kit Urine protein analysis can be used for the identification of potential biomarkers in urine, and to diagnose and/or monitor renal and other diseases. Due to the fact that urine can be collected non-invasively in large amounts, it provides an attractive alternative to blood plasma as a potential source of disease biomarkers. The ProteoSpin Urine Protein Concentration Micro Kit provides a fast and simple procedure for concentrating dilute solutions of urine proteins from small volumes of urine. The kit employs spincolumn chromatography using Norgen s proprietary protein resin as an ion exchanger. Urine proteins will bind to the column, while nonspecifically bound proteins and salts are easily removed from the sample. Each spin column is able to concentrate and desalt up to 200 µg of urine proteins. The simultaneous removal of salts while concentrating a dilute urine protein solution makes the kit a convenient method for preparing urine proteins before running downstream proteomic applications including SDS-PAGE, 2D gels, whole protein mass spectrometry, and protein microarrays. ProteoSpin Urine Protein Concentration Micro Kit Benefits Fast and easy processing Complete kit No molecular weight cutoff Proteins bind while salts are discarded in flowthrough Suitable for downstream applications Efficiently process up to 12 samples in only 20 minutes using an easy-to-use protocol. All columns and solutions required are provided. Based on an ion exchange mechanism, a broad size range of proteins can be processed. Norgen s resin has an inherent low affinity for salt, and high affinity for proteins - providing salt removal and protein concentration simultaneously. Final elution suitable for: SDS-PAGE 2D gel electrophoresis Whole protein mass spectrometry Protein microarrays

31 A B C A B C D E F Figure 1. When using the ProteoSpin Urine Protein Concentration Kit, no urine proteins are lost during the binding step, as can be seen by examining the binding flowthrough. Lane A is input urine, Lane B is the binding flowthrough, and Lane C is the elution. Figure 2. A dose response indicating the concentration of urine proteins from increasing input amounts of urine. Lane A represents 25 µl of an input urine sample, while lanes B to F are the elutions after the proteins were concentrated using the ProteoSpin Urine Protein Concentration Micro Kit. The input amounts of urine that were loaded onto each column were 100 µl, 200 µl, 300 µl, 400 µl, and 500 µl respectively. Urine Protein Concentration Micro Kit Contents Each Urine Protein Concentration Micro Kit provides essential components for use with the provided protocols. 1. ph Binding Buffer 2. Column Activation and Wash Buffer 3. Protease Inhibitor Mix 4. Stabilizer 5. Elution Buffer 6. Neutralizer 7. Micro Spin Columns 8. Elution Tubes 9. Short Protocol Card Customer-Supplied Reagents and Equipment Benchtop microcentrifuge Micropipettors Cat # Description Quantity ProteoSpin Urine Protein Concentration Micro Kit 25 samples

32 ProteoSpin Serum Depletion Kit The ProteoSpin Serum Depletion Kit provides a fast and simple procedure for the effective depletion of major serum proteins including albumin, α-antitrypsin, transferrin and haptoglobin from serum and plasma samples. The kit is unique in that it is based on an ion-exchange mechanism and not on the use of specific antibodies. As a result, the kit can be used to deplete serum proteins from a wide variety of samples, including human and various animals. Each spin column can deplete up to 500 µg of serum proteins. Albumin has been found to be depleted by 70%, transferrin and haptoglobin by 50% and α-antitrypsin by 90%. The complexity of the sample is thus greatly reduced, allowing for the detection of less abundant proteins present in the sample. Eluted samples are ready for use in various downstream applications including 2D gel electrophoresis, LC/MS and microarrays. ProteoSpin Serum Depletion Kit Benefits Efficient removal of highly abundant proteins Fast and easy processing Does not rely on antibodies Complete kit Low price per column Depletion of proteins and salts Suitable for downstream applications The kit depletes 70% of albumin, 90% of α-antitrypsin, and 50% of transferrin and haptoglobin, enabling the visualization of low abundance proteins. Efficiently process up to 10 samples in less than 30 minutes using an easy-to-use protocol. Based on an ion exchange mechanism, the kit can be applied to serum and plasma samples from a range of sources, including human and various animals. All columns and solutions required are provided. Compared to columns based on antibodies, these ion-exchange columns are much lower in price, yet perform to the same high standards. Most proteins bind to the column, while salts and many of the major proteins are removed in the flowthrough. Final elution suitable for: SDS-PAGE 2D gel electrophoresis LC/MS Microarrays

33 Figure 1. Abundant proteins are depleted from human serum to a large degree using the ProteoSpin Serum Depletion Kit. Lane A represents the input human serum proteins, while Lanes C and D represent the elution after the abundant proteins were depleted with the kit. Lane B represents the binding flowthrough, and contains the proteins that have been depleted from the serum sample. Figure 2. A 2D gel was run after the abundant serum proteins were depleted from human serum using the ProteoSpin Serum Depletion Kit. The abundant proteins including albumin, α- antitrypsin, transferrin and haptoglobin have been removed from the sample, allowing the visualization and detection of the lower abundance proteins of interest. Serum Depletion Kit Contents Each Serum Depletion Kit provides essential components for use with the provided protocols. 1. Column Activation and Wash Buffer 2. Elution Buffer 3. Neutralizer 4. Mini Spin Columns 5. Elution Tubes 6. Short Protocol Card 7. Application Manual Customer-Supplied Reagents and Equipment Benchtop microcentrifuge Micropipettors Cat # Description Quantity ProteoSpin Serum Depletion Kit 25 samples

34 ProteoLadder 100, ProteoLadder 125, ProteoLadder 150 Applications: ProteoLadder protein markers are ideal for accurate molecular weight sizing of proteins by SDS-PAGE gels stained with Coomassie blue. Features: Available in ready-to-use and lyophilized formats Highly stable Lyophilized ladders are easy to reconstitute Optimized storage solution to ensure ladder stability even after multiple freeze/thaws Higher intensity reference bands Flexible molecular weight choices; three products ranging from 20 kda to 150 kda ProteoLadder 100 ProteoLadder 125 ProteoLadder 150 Contents: Ready to use: 1 ml premixed ladder in loading buffer (50 mm Tris HCl, ph 7.5, 100 mm DTT, 0.01% bromophenol blue, 1 mm EDTA, 1 mm sodium azide, 2% SDS, 20% glycerol). Lyophilized: 1 vial lyophilized ladder protein, 1 vial 600 µl loading dye (50mM Tris HCl, ph 7.5, 100 mm DTT, 0.01% bromophenol blue, 1 mm EDTA, 1 mm Sodium Azide, 2% SDS, 20% glycerol). Cat # Description Quantity ProteoLadder 100, Lyophilized 100 loads ProteoLadder 100, Ready-to-use 100 loads ProteoLadder 125, Lyophilized 100 loads ProteoLadder 125, Ready-to-use 100 loads ProteoLadder 150, Lyophilized 100 loads ProteoLadder 150, Ready-to-use 100 loads

35 Corporate Headquarters 344 Merritt St. St. Catharines, Ontario CANADA L2T 1K6 Phone: (905) or Toll Free in North America: Fax: (905) Ordering Information Norgen Biotek offers several ways to place orders 24 hours a day. Phone: (905) or Toll Free in North America: Website: orders@norgenbiotek.com Fax: (905) To locate a distributor in your area, please contact Norgen Biotek. Technical Support Contact our Technical Support Team between the hours of 8:30 and 5:30 (Eastern Standard Time) at (905) or Toll Free at Technical support can also be obtained from our website ( or through at techsupport@norgenbiotek.com. Trademarks and Disclaimers Norgen s protein purification technology is covered by the pending U.S. patents 10/294,283 and 60/331464, as well as the international PCT patent application PCTCA ProteoSpin is a registered trademark of Norgen Biotek Corporation. Some product compositions are proprietary and undisclosed. The following trademarks also used in this publication are the property of the companies listed below. Triton is a registered trademark of Rohm & Haas Co. ZipTip is a trademark of Millipore Corporation.

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