TECHNOLOGY RunBlue TM Gels PROTEINS ELECTROPHORESIS

Transcription

1 TECHNOLOGY RunBlue TM Gels RunBlue TM precast gels have been developed as an improvement on the current state of the art precast SDS-PAGE gels. RunBlue TM gels are more rigid and up to 10 times stronger than conventional precast gels, resulting in more robust gels that can be handled with confidence without tearing - an 8% gel can be thrown into the air and caught without breaking! BENEFITS OF RUNBLUE TM PRECAST GELS PROTEINS ELECTROPHORESIS Mass Spec compatible 2 Year shelf life - never throw away another gel Up to 10x stronger - no more torn, ruined gels No comb to remove - no more broken or bent teeth Full length resolving gel - improved resolution and better blotting Fast delivery with immediate and expert technical support for total peace of mind Best value gels available - save money! Proprietary photo-polymerisation process Composite Gel technology Unique plastic cassette Unique proprietary stacking chemistry Optimised Running buffer RUNBLUE TM TECHNOLOGY - Sharper bands - Better batch to batch reproducibility - Stronger gels - Reduced hydrophobicity -> less streaking - Enhanced oxygen barrier -> sharper bands - Better stacking -> sharper bands - Larger volume capacity - Lower conductivity -> Less heat generation - Run at higher voltage -> shorter run 28

2 RunBlue TM gels are packed with innovative technology focused on delivering the highest resolution and band sharpness. A direct comparison between RunBlue TM and a leading competitor product demonstrates the superior band sharpness and resolution that can be obtained with RunBlue TM gels. Competitor 4-12% RunBlue TM 4-20% Lane 1: Biorad Marker. Lane 2: BSA. Lane 3: Chicken Muscle Lysate. Lane 4: E. Coli Lysate. Samples were prepared in LDS sample buffer and gels were run at 200V in appropriate running buffers. RUNBLUE TM SPECIFICATIONS Gel percentages 8%, 10%, 12%, 16%, 4-8%, 4-12%, 4-20% Storage & Shelf Life Pack size Running buffer Sample buffer WxHxD (cm) Sample volume 24 months at 2º - 8ºC (DO NOT FREEZE) 3 months at room temperature 10 gels per box TEA-Tricine-SDS (NXB50500), TEO-Tricine Native (NXB61500), Bis-Tris MES (NXB70500), Bis-Tris MOPS (NXB75500) LDS for TEO-Tricine (NXB31010), LDS for Bis-Tris (NXB32010) Cassette dimensions 10 x 10 - NXG, NBT 8 x 10 - BCG Gel dimensions VIRTUALLY IDENTICAL RESULTS AFTER ONE YEAR 10 x 10 x x 10 x x 8 x x 8 x well 700 µl 700 µl 2 well 450 µl 450 µl 12 well 35 µl 35 µl 17 well 20 µl 20 µl 2 Weeks Old 1 Year Old RunBlue TM gels are extremely strong and durable. Not only is the product 10x stronger than any competitor product, RunBlue TM gels also have an unsurpassed two year shelf life. expedeon.com 29 DNA RNA ELECTROPHORESIS WESTERN BLOTTING QUANTIFICATION PREPARATION PURIFICATION FRACTIONATION SPECTROSCOPY NUCLEIC ACIDS PROTEINS

3 Bis-Tris Protein Gels RUNBLUE TM RunBlue TM Bis-Tris Protein Gels have been developed by Expedeon to provide an alternative to NuPAGE Bis-Tris gels. RunBlue TM Bis-Tris Protein Gels run with the same voltages and times, in the same tanks, using the same MES/MOPS buffers as NuPAGE Bis-Tris gels and achieve highly comparable results with exceptional reproducibility. RunBlue TM Bis-Tris Protein Gels also benefit from the usability enhancements present in all RunBlue TM gels such as comb/strip free design and tear proof composition. Comparable Results to NuPAGE Bis-Tris Gels Same protocols, buffers and gel tank PROTEINS ELECTROPHORESIS KEY BENEFITS DON T CHANGE A THING More practical comb free, strip free, tear proof gels Standard Bis-Tris Buffers Compatible with MES and MOPS buffers Standard format Compatible with NuPAGE XCell SureLock Mini-Cell and similar tanks Standard Bis-Tris Protocol Fast run times at 200V Neutral running ph Sharp bands and high protein integrity Unique homogeneous polymerisation Increased consistency and no residual free acrylamide Extra Wells Run up to 12 or 17 samples simultaneously Deep and wide wells Load up to 35 μl (12 well) or 20 μl (17 well) Protruding teeth No well contamination in case of overloading Resilient gel matrix Tear-proof gel and risk-free handling Comb-free Reduced risk of broken or damaged teeth Strip-free No potentially ruined Western Blots Neutral buffer and high chemical stability Shelf life: 1 year at 4 C and 3 months at Room Temp. Expedeon s new Bis-Tris Gels can be run with standard MES or MOPS buffers depending on the molecular weight range of proteins to be separated and use the same LDS sample buffer and protocols as NuPAGE (200V). The cassettes fit in the NuPAGE SureLock TM tank without any need for adaptors or spacers, and gels are also compatible with the iblot dry blotting system. There is no workflow disruption, no downtime, and no transition period. 30

4 USABILITY ENHANCEMENTS In addition to their high resolution separation of protein all RunBlue TM Protein Gels have a number of enhancements to improve and streamline usability. RunBlue TM Protein Gels are up to 10 times stronger than conventional hand-cast gels and can stand up to repeated handling. This practically eliminates the risk of torn and unusable gels encountered during normal gel manipulation. More lanes than standard gels allows more proteins to be run and wide, deep wells allow higher volumes to be loaded. Cassettes have no comb or tape to remove and no excess gel to trim, all of which improves the speed and efficiency of the loading and running process. Wells are outlined and numbered for quicker sample loading, and each cassette is labelled with a unique lot number for individual traceability. Well teeth protrude above the cassette to prevent sample contamination between wells in case of overloading. DON T COMPROMISE ON RESULTS Expedeon have over 10 years experience in pre-cast gel manufacture and have applied this knowledge to Bis-Tris gel chemistry. A unique manufacturing process with exacting specifications and quality controls results in increased batch to batch consistency between gels. A key element in this process is the use of physically induced gel polymerisation for consistency, as oppose to chemically induced methods which can introduce variability leading to heterogeneous polymerisation. Switching to RunBlue TM Bis-Tris gels will give better resolution while following the same migration profiles. Code NXB70500 NXB75500 NXB % 20% 40% 60% 80% 100% Description Bis-Tris 8% 8% 10% 10% 12% 12% 4-12% 4-12% MES MOPS MES MOPS MES MOPS MES MOPS Each box contains 10 cassettes GEL PERCENTAGE 12 WELL 17 WELL 10 x 10 cm cassette Compatible with SureLock TM tanks MES Run Buffer - 20x ml MOPS Run Buffer - 20x RunBlue TM LDS Sample Buffer - 4x % NBT00812 NBT % NBT01012 NBT % NBT01212 NBT % (Gradient) NBT41212 NBT41227 expedeon.com 31 DNA RNA ELECTROPHORESIS WESTERN BLOTTING QUANTIFICATION PREPARATION PURIFICATION FRACTIONATION SPECTROSCOPY NUCLEIC ACIDS PROTEINS

5 Teo-Tricine SDS & Native Gels RUNBLUE TM RunBlue TM SDS Protein Gels take advantage of innovations to the gel making process to increase resolution and reproducibility of separated protein bands. RunBlue TM SDS Protein Gels are based on a TEO-Tricine gel buffer system which provide good separation profiles with enhanced separation of higher molecular weight proteins and better overall resolution. RunBlue TM SDS protein gels are designed for high resolution SDS-PAGE. Their novel chemistry system and unique manufacturing process ensure very sharp bands and reproducible results. They have all the usability enhancements associated with RunBlue TM gels, such as being tear proof and comb free, and are compatible with a number of gel running tanks. RunBlue TM SDS Protein Gels are optimised for a TEO-Tricine buffer system and should be run with the Tricine SDS Run buffer. PROTEINS ELECTROPHORESIS SDS NATIVE 10 x 10 cm Cassette Compatible with SureLock Tanks 8 x 10 cm Cassette Compatible with BioRad Tanks 10 x 10 cm Cassette Compatible with SureLock Tanks 8 x 10 cm cassette Compatible with BioRad Tanks Gradient Single % Gradient Single % Gradient Single % Gradient Single % 1 Well 2 Well 12 Well 17 Well 8% NXG00801 NXG00802 NXG00812 NXG % NXG01001 NXG01002 NXG01012 NXG % NXG01201 NXG01202 NXG01212 NXG % NXG01601 NXG01602 NXG01612 NXG % NXG40801 NXG40802 NXG40812 NXG % NXG41201 NXG41202 NXG41212 NXG % NXG42001 NXG42002 NXG42012 NXG % BCG00812 BCG % BCG01012 BCG % BCG01212 BCG % BCG01612 BCG % BCG40812 BCG % BCG41212 BCG % BCG42012 BCG % NXN01012 NXN % NXN02012 NXN % NXN20812 NXN % NXN32012 NXN % BCN01012 BCN % BCN02012 BCN % BCN20812 BCN % BCN32012 BCN

6 PROTEIN MIGRATION CHARTS RunBlue TM Migration Chart: 10x10 cm TRICINE Running buffer (NXB50500) RunBlue TM Migration Chart: 8x10 cm TRICINE Running buffer (NXB50500) COMPATIBLE TANKS GEL BOX MFG COMPATIBLE BOXES Mini-Protean II, Mini-Protean III 8 x 10 cm Bio - Rad Mini-Protean Tetra Hoefer Hoefer TM TallMighty Small TM (SE280) 10 x 10 cm Life-Technologies X-cell TM Bolt Lonza PAGEr Minigel Chamber expedeon.com 33 DNA RNA ELECTROPHORESIS WESTERN BLOTTING QUANTIFICATION PREPARATION PURIFICATION FRACTIONATION SPECTROSCOPY NUCLEIC ACIDS PROTEINS

7 TEO-TRICINE SDS GELS CONVERSION TABLES GEL CONVERSION TABLE: LIFE NUPAGE --> RUNBLUE TM NuPage Bis Tris RunBlue TM TEO-CI Gel Type Running Buffer Gel Type Running Buffer 8% 8% or 4-8% 10% 10% MOPS 12% 12% TEO-Tricine (NXB50500) 4-12% 4-12% 8% 10% 10% 12% MES 12% 16% TEO-Tricine (NXB50500) 4-12% 4-20% GEL CONVERSION TABLE - WELL TYPE: LIFE NUPAGE --> RUNBLUE TM NuPage Bis Tris RunBlue TM TEO-CI Well Type Volume (µl) Well Type Volume (µl) PROTEINS ELECTROPHORESIS Biorad TGX GEL CONVERSION TABLE: BIORAD TGX --> RUNBLUE TM RunBlue TM TEO-CI Gel Type Running Buffer Gel Type Running Buffer 7.5% 8% 10% 10% 12% 12% 4-15% Tris Glycine 4-12% 4-20% 4-20% 8-16% 12%-16% Any Kda 4-20% GEL CONVERSION TABLE - WELL TYPE: BIORAD TGX --> RUNBLUE TM Biorad TGX RunBlue TM TEO-CI TEO-Tricine (NBX50500) Well Type Volume (µl) Well Type Volume (µl)

8 Premixed Buffers & Solutions RUNBLUE TM All RunBlue TM Buffers, Reagents and Accessories have been specifically designed and formulated for use with the RunBlue TM gels and apparatus to provide optimum performance and results. Buffers are made using the highest quality chemicals and have different compositions to other manufacturers buffers. Only use RunBlue TM formulated buffers with RunBlue TM gels. RunBlue TM LDS Sample Buffer (NXB31010) has been specifically formulated for use with RunBlue TM gels. The ions in the sample buffer match the gel buffer and the buffer has a higher density, making it compatible with the density of the running buffer. The buffer contains two tracking dyes. The red dye is designed to monitor the ph of the buffer. A colour change from red to yellow indicates a shift in ph indicating that the buffer was poorly made or recycled too many times. RunBlue TM SDS Running Buffer (NXB50500) is a TEO-Tricine buffer system that can be used under reducing or non-reducing conditions. TEO-Tricine running buffer can be used for excellent separation of higher or lower molecular weight proteins. SDS NATIVE BLOTTING Code Description Unit Size NXB50500 RunBlue TM Tricine - SDS Run Buffer 20X 500 ml NXB50425 RunBlue TM Tricine - SDS Run Buffer 20X 4 L NXB31010 RunBlue TM LDS Sample Buffer 4X 10 ml NXA30010 RunBlue TM Antioxidant 10X 10 ml NXA32001 RunBlue TM Sample Reducer (DTT) 10X 1 ml NXB70500 RunBlue TM MES Run Buffer ml (20x Concentrate) 500 ml NXB70425 RunBlue TM MES Run Buffer - 4 L (20x Concentrate) 4 L NXB75500 RunBlue TM MOPS Run Buffer ml (20x Concentrate) 500 ml NXB75425 RunBlue TM MOPS Run Buffer - 4 L (20x Concentrate) 4 L NXB61500 RunBlue TM Native Run Buffer 20X 500 ml NXB61425 RunBlue TM Native Run Buffer 20X 4 L NXB33010 RunBlue TM NATIVE Sample Buffer 4X 10 ml NXB86500 RunBlue TM Tris Glycine Blot Buffer 10X 500 ml NXB86425 RunBlue TM Tris Glycine Blot Buffer 10X 4 L NXB82500 RunBlue TM Tris Glycine SDS Buffer 10X 500 ml NXB82425 RunBlue TM Tris Glycine SDS Buffer 10X 4 L NXA19020 RunBlue TM Blot sandwich Nitrocellulose membrane (90x85 mm) µm pack of 20 NXA29320 RunBlue TM Blot sandwich PVDF membrane (90x85 mm) µm pack of 20 expedeon.com 35 DNA RNA ELECTROPHORESIS WESTERN BLOTTING QUANTIFICATION PREPARATION PURIFICATION FRACTIONATION SPECTROSCOPY NUCLEIC ACIDS PROTEINS

9 Protein Markers RUNBLUE TM RunBlue TM prestained markers enable easy visualization of the marker proteins during and after running and western blotting. The makers can be used to estimate the molecular weight of your protein of interest and assess the transfer efficiency during bloting. Expedeon offers two types of prestained molecular weight markers. MARKER TRI COLOUR DUAL COLOUR Range 180 KDa - 5 KDa KDa kda 115 # of bands Blue 180, 144,115, 92, 56, 40, 27, 14, 12, Blue 195, 142, 96, 48, 33, 22, 12, PROTEINS ELECTROPHORESIS Colours Storage RECOMMEND LOAD VOLUMES RunBlue TM prestained markers are provided ready to use in 1x RunBlue TM sample buffer. Recommended volumes to load are: APPLICATION Red Pink 71, Green 18 RunBlue TM SDS 4-20% Gel 12 WELL 17 WELL VOLUME LOADS / VIAL VOLUME LOADS / VIAL Monitor during run 10 µl 60 5 µl 120 Visible after blotting 4 µl µl 300 Visible after staining 4 µl µl 300 for use with Antiblue 2 µl µl 600 RunBlue TM SDS 4-20% Gel Storage long term at -20ºC or for 3 months at 4ºC. Code Description Unit Size NXA05160 RunBlue TM Dual Colour marker 600 µl NXA6050 RunBlue TM Tri Colour marker 500 µl RunBlue TM Tri Colour marker run on different gel chemistries 36

10 Equipment & Accessories RUNBLUE TM RunBlue TM gels run best on our Dual Run and Blot unit. This unique apparatus permits the running of both 10x10cm and 8x10cm gels. The unit is available in dual and quad form, enabling up to 4 gels to be run at one time. Both units can also be used to complete Western blots. The unique electrode arrangement provides a gradient electric field which ensures that both small and large proteins migrate from the gel at a comparable rate. Quad units are available in cooled format, enabling cold water hook up if required. Code Gel Tanks NXE00002 NXE00004 NXE00005 NXE00014 NXE00015 NXE00016 NXE00017 Blot Membrane Fits both 10x10 and 8x10 cassettes Coloured centre core to provide maximum contrast with loading buffer for easy loading Excellent sealing and no leakage from cathode to anode Can be cooled in place to reduce heat effects such as smiling or frowning Description RunBlue TM Dual Run and Blot System (runs up to 2 gels) supplied with 1 bufferdam, 2 blotting cassettes and 2 cool blocks RunBlue TM Quad Run and Blot System (runs up to 4 gels) supplied with 1 bufferdam and 4 blotting cassettes RunBlue TM Quad Cool (runs up to 4 gels) supplied with 1 bufferdam and 4 blotting cassettes RunBlue TM Blotting Cassette with Pad 9x9 cm RunBlue TM Cooling Block, Dual Run and Blot RunBlue TM Buffer Dam, Dual Run and Blot RunBlue TM Sponge Pad Kit, 9x9 cm, 4 pack NXA19020 RunBlue TM Blot Sandwich NC 90x85mm, 1 pack of 20 NXA29320 RunBlue TM Blot Sandwich PVDF 90x85mm, 1 pack of 20 Gradient field for blotting providing more uniform transfer. Ideal for high quality wet blotting Excellent value for money RunBlue TM Tanks last 10x longer than SureLock Tank RunBlue TM Tanks run gels evenly compared to the SureLock Tanks RunBlue TM Tanks can run and blot expedeon.com 37 DNA RNA ELECTROPHORESIS WESTERN BLOTTING QUANTIFICATION PREPARATION PURIFICATION FRACTIONATION SPECTROSCOPY NUCLEIC ACIDS PROTEINS

11 Acrylamide RUNBLUE TM RunBlue TM ready to use acrylamide / bis-acrylamide solutions present a quicker, safer, more reliable way to hand cast gels for PAGE without having to use powdered acrylamide. RunBlue TM acrylamide solutions come in a variety of volumes, percentages, and common cross-linker ratios in a pre-filtered, ready to use format to ensure quality and reproducibility between gels. BENEFITS 40% Acrylamide solution For making SDS PAGE gels For use with Expedeon Gel casting equipment Supplied in volumes of 500ml / 100ml Multiple filtering rounds to ensure quality Provide as solution, so no dust inhalation risk and reduced contact hazard Polymerise with APS and TEMED RunBlue TM acrylamide is available in 100ml and 500ml volumes at 30% and 40% stock solutions. For the full range of solution percentages and ratios of acrylamide / bis-acrylamide solutions, see the table below: SOLUTION VOLUME ACRYLAMIDE : BIS-ACRYLAMIDE SUGGESTED USE 19:1 (5% crosslinker) Peptide / DNA separation PROTEINS ELECTROPHORESIS 30% 100ml or 500 ml 29:1 (3.3% crosslinker) Peptide / DNA separation 37:1 (2.7% crosslinker) Higher MW protein separation 19:1 (5% crosslinker) Peptide / DNA separation 40% 100 ml or 500 ml 29:1 (3.3% crosslinker) Peptide / DNA separation 37:1 (2.7% crosslinker) Higher MW protein separation Use with RunBlue TM empty cassettes (8x10cm or 10x10cm) and combs (12 or 17 well) to produce reliable hand cast gels. Code Description A Acrylamide Solution 40% for electrophoresis - sterile filtered ml A Acrylamide Solution 40% for electrophoresis - sterile filtered ml A Acrylamide Solution 30% for electrophoresis - sterile filtered ml A Acrylamide Solution 30% for electrophoresis - sterile filtered ml A Acrylamide/Bis-acrylamide 40% solution 19:1-100 ml A Acrylamide/Bis-acrylamide 40% solution 19:1-500 ml A Acrylamide/Bis-acrylamide 30% solution 19:1-100 ml A Acrylamide/Bis-acrylamide 30% solution 19:1-500 ml A Acrylamide/Bis-acrylamide 40% solution 29:1-100 ml A Acrylamide/Bis-acrylamide 40% solution 29:1-500 ml A Acrylamide/Bis-acrylamide 30% solution 29:1-100 ml A Acrylamide/Bis-acrylamide 30% solution 29:1-500 ml A Acrylamide/Bis-acrylamide 40% solution 37:1-100 ml A Acrylamide/Bis-acrylamide 40% solution 37:1-500 ml A Acrylamide/Bis-acrylamide 30% solution 37:1-100 ml A Acrylamide/Bis-acrylamide 30% solution 37:1-500 ml 38

12 Empty Cassettes ACCESSORIES Expedeon empty cassettes are exactly the same cassettes as used in the RunBlue TM precast gel range. After many years of producing high quality precast gels Expedeon cassettes have been adapted for self-casting to permit easy production of high quality gels as required, at a much more economical price point than other manufacturer s empty cassettes and combs. Expedeon cassettes have been developed over many years to combine strength, ease of use, and reliability in one package. RunBlue TM cassettes are pre-taped to save time when pouring gels, and help to avoid the common errors that can be encountered when casting with glass plates. Combs are available in 12 or 17 well formats. Cast gels are 1mm thick when set. Expedeon cassettes are available in 10x10cm and 8x10cm formats and 12 or 17 well combs are available to strike a balance between resolution, sample loading volume and number of samples per gel. Combs are compatible with both 8x10cm and 10x10cm cassettes. RunBlue TM cassettes help to make reliable self-cast gels easy to produce. Code BCC01010 BCC00810 BCC00012 BCC00017 Description RunBlue TM Empty Cassettes (pack of 25) 10x10cm 1.0mm RunBlue TM Empty Cassettes (pack of 25) 8x10cm 1.0mm RunBlue TM Cassette Comb (pack of 25) - 12 Well RunBlue TM Cassette Comb (pack of 25) - 17 Well BENEFITS Extensively tested cassettes Pre-taped 8x10 cm, 10x10 cm, 12 or 17 well More convenient than glass plates Expedeon also provides RunBlue TM acrylamide, a ready to use pre-filtered acrylamide/bis-acrylamide solution for gel casting. expedeon.com 39 DNA RNA ELECTROPHORESIS WESTERN BLOTTING QUANTIFICATION PREPARATION PURIFICATION FRACTIONATION SPECTROSCOPY NUCLEIC ACIDS PROTEINS

13 Stain Tray ACCESSORIES The Expedeon Stain Tray is a lightweight, simple solution for staining gels with InstantBlue TM or any other protein stain. Trays are large enough for midi gels, stackable, and come with a lid to help avoid spills and allow stacking. PROTEINS ELECTROPHORESIS Lightweight design Easy removal lid design with protruding tabs to separate lid and tray Lid protects from evaporation and contamination Can be stacked on rockers/shakers or in fridge Pour spouts in each corner of tray Transparent plastic to allow staining progress checks at a glance RNase, DNase and edontoxin free Reusable Expedeon Staining Trays are designed to provide a cost effective, reusable staining vessel. Pour spouts in every corner reduce mess and waste if reusing or disposing of stain. Tightly sealing lids prevent evaporation when storing gels and allow stacking on rockers and in fridges. Both lids and trays are reusable, and when not in use trays and lids stack separately to minimise storage space required. Trays are made from APET and are not microwavable. Dimensions (Internal): 10.0 cm x 14.3 cm x 3.0 cm. Code ISBST Description Stain Tray - InstantBlue TM 40

14 Expedeon products can be ordered from: