Protein & Antibody. Purification & Detection Tools

Transcription

1 Abbkine featured PurKine isolation and purification portfolio with complete antitag antibodies and synthetic peptides to meet and satisfy your most types of protein purification, sample preparation and detection analysis. Protein & Antibody Purification & Detection Tools

2 PRODUCT TABLE INTRODUCTION Protein Purification & Detection His Tag Fusions GST Tag Fusions MBP Tag Fusions DDDDK Tag Fusions Biotin Tag Fusions HA Tag Fusions Protein A Tag Fusions Myc Tag Fusions VSV-G Tag Fusions Other Tag Fusions Antibody Purification & Detection Protein A Protein G Protein L Protein A/G Antibody Purification IPKine Antibodies IFKine Antibodies Proteins are large, complex molecules that play many critical roles in the body. They do most of the work in cells and are required for the structure, function, and regulation of the body s tissues and organs. Protein purification is a fundamental step for analyzing individual proteins and protein complexes and identifying interactions with other proteins, nucleic acids. Because purification of native proteins can be challenging, affinity purification tags, including His Tag, GST Tag, S Tag, HA Tag, DDDDK Tag, Myc Tag, MBP Tag and Biotin Tag are often fused to a recombinant protein of interest such that the tag is used to capture or detect the protein. Many methods have been used to capture or purify a protein of interest from crude extracts or other complex mixtures. The most powerful of these methods is affinity chromatography, whereby the protein of interest is purified by virtue of its specific binding properties to an immobilized ligand. Most significantly, the well-characterized tagligand chemistry enables single-step affinity purification of tagged molecules using immobilized versions of their corresponding ligands. Antibodies to fusion tags are also widely available for use in downstream detection and assay methods, eliminating the need to obtain or develop a probe for each specific recombinant protein. Antibody purification involves selective enrichment or specific isolation of antibodies from serum (polyclonal antibodies), ascites fluid or cell culture supernatant of a hybridoma cell line (monoclonal antibodies). Protein A, Protein G and Protein L are three bacterial proteins whose antibody-binding properties have been well characterized. These proteins have been produced recombinantly and used routinely for affinity purification of key antibody types from a variety of species. Purification of antigen-specific antibodies is also required for some cases. Abbkine offer featured PurKine affinity chromatography portfolio with complete Tag monoclonal antibodies and synthetic peptides to meet and satisfy your most types of protein purification, sample preparation and detection analysis.

3 His Tag Fusions - Purification, Detection and Blocking Polyhistidine-tag is an amino acid motif in proteins that consists of at least six histidine (His) residues, often at the N- or C-terminus of the protein. It is also known as hexa histidine-tag, 6xHis-tag, His6 tag and His-tag. His-tag are often used for affinity purification of His-tagged recombinant proteins expressed in Escherichia coli and other prokaryotic expression systems. His-tag fusion proteins can be purified and detected easily because the string of histidine residues binds to several types of immobilized metal ions, including nickel, cobalt and copper, under specific buffer conditions. In addition, anti-his-tag antibodies are commercially available for use in assay methods involving Histagged proteins. His-Tag Protein Purification Tools Effetive purification of His tag fusion proteins with NTA, IMCA, IDA formated resins and columns His Tag Fusion - Purification, Detection and Blocking Immobilized metal affnity chromatography (IMAC) is a popular method for protein purifcation, particularly for recombinant proteins fused to a polyhistidine tag. Ni-NTA resins have been the most common IMAC resin choice for 6xHis-tag protein purifications because of the four metal-binding sites on the chelate, which enables high-protein binding and low-metal ion leaching.you can also choose other metal chelating, like Cu2+ or Co2+ ligand products or just chelate by yourself in case you re familiar with protein purification procedure. Fig. 1 A B C PurKine His-Tag Purification system is based on innovative high-capacity IMAC matrix for convenient single-step purification of His-tag proteins from total lysates. Our propriety ion-chelate chemistry ensures extraordinary compatibility with commonly used reducing agents such as DTT, chelating metalloprotease inhibitors such as EDTA, and a wide range of buffer substances and salt conditions. The portfolio provides wide choice of chelating group (IDA, NTA or Super NTA), cross-linked agarose (crosslinked 4% or 6% agarose) or magarose, ions (Nick, Colbat, Copper, or non ion charged) and compatible ingredients to allow optimization of process for maximum protein yield, stability and solubility. Fig. 1. His recombinant proteins were purified with PurKine His- Tag Ni-NTA Resin 4FF (Lane B) and counterpart product from supplier G (Lane C). PurKine His-Tag Purification resin is also available as prepacked spin column and kit formats. Agarose resin and magbeads formation are both available. PurKine His-Tag Ni-IDA Resin Agarose Resin BMR ml, 10ml, 50ml, 100ml PurKine His-Tag Ni-NTA Resin Agarose Resin BMR ml, 10ml, 50ml, 100ml PurKine His-Tag Ni-NTA Packed Column Packed Column BMC ml 5, 3ml 5 PurKine His-Tag Protein Purification Kit (Ni-NTA) Kit (Agarose Resin) KTP ml 5, 3ml 5 PurKine His-Tag Ni-NTA Resin 6FF Agarose Resin BMR ml, 10ml, 50ml, 100ml PurKine His-Tag Ni-NTA Packed Column 6FF Packed Column BMC ml 5, 3ml 5 PurKine His-Tag Cu-IDA Resin Agarose Resin BMR ml, 10ml, 50ml, 100ml PurKine His-Tag Co-NTA Resin Agarose Resin BMR ml, 10ml, 50ml, 100ml PurKine His-Tag Co-NTA Resin 6FF Agarose Resin BMR ml, 10ml, 50ml, 100ml PurKine His-Tag IMAC-NTA Resin 6FF Agarose Resin BMR ml, 10ml, 50ml, 100ml PurKine His-Tag Ni-NTA Magbeads Agarose Magbeads BMM ml, 5ml, 10ml 1

4 His Tag Fusion - Purification, Detection and Blocking PurKine His-Tag Ni-Super Resin is a superior replacement to Ni-NTA Resin, which has been charged with nickel ions (Ni2+) more closely thus have more stable binding with Ni2+ than NTA, thus featured more resistance to various chemicals including reducing, chelating agents. PurKine His-Tag Ni-Super Resin is suitable for purification of His-tag proteins from samples that can cause Ni stripping from the medium, e.g. secreted proteins in solution containing chelators. In addition, its high flow properties make it excellent for scaling-up. Fig. 2 Fig. 3 A A B C C D D E E A F B G H C D I EJ Fig. 2. PurKine His-Tag Ni-Super Resin featured more resistance to various chemicals such as 0.01M HCl (Lane C) and 0.01M NaCl (Lane D). Fig. 3. PurKine His-Tag Ni-Super Resin featured more resistance to various chemicals such as 6M Gua-HCl (Lane D) and 1M NaCl (Lane E) after 24hr elution. PurKine His-Tag Ni-Super Resin Resin BMR ml, 10ml, 50ml, 100ml PurKine His-Tag Ni-Super Packed Column Packed Column BMC ml 5, 3ml 5 PurKine His-Tag Ni-Super Resin 6FF Resin BMR ml, 10ml, 50ml, 100ml His-Tag Protein Detection Antibodies Specific detection of His Tag fusion proteins with His tag monoclonal and polyclonal antibodies Anti-His tag antibodies are widely used to monitor the progress of purification by western blotting. Antibodies are also handy in order to ensure no proteolytic degradation of the recombinant protein is taking place. Anti-His Tag Mouse Monoclonal Antibody (5C3) WB, IF, IP A μl, 200μl, 1ml, 10ml Anti-His Tag Rabbit Polyclonal Antibody WB A μl, 200μl, 1ml, 10ml HRP Conjugated Anti-His Tag Mouse Mab (5C3) WB A μl, 200μl, 1ml, 10ml His-Tag Blocking Peptide High quality blocking peptide for Anti-6X His tag antibody Hexa His Tag Peptide is used for blocking the antibody activity of His tag antibody. It usually blocks the antibody activity completely in Western blot analysis by incubating the peptide with equal volume of antibody. Hexa His Tag Peptide WB, BL PRQ mg, 20mg, 100mg 2

5 GST Tag Fusions - Purification and Detection Glutathione S-transferase (GST) is a 211 amino acid protein (26kDa), which can be fused to either the N-terminus or C-terminus of a protein. Because GST rapidly folds into a stable and highly soluble protein upon translation, inclusion of the GST tag often promotes greater expression and solubility of recombinant proteins than expression without the tag. GST-tagged fusion proteins can be purified or detected based on the ability of GST (an enzyme) to bind its substrate, glutathione (GSH). GST-Tag Protein Purification Tools Effetive purification of GST tag or GST fusion proteins with agarose resins or magbeads PurKine GST-Tag Purification system is based on innovative high-capacity matrix for convenient single-step purification of recombinant GST fusion proteins and other glutathione binding proteins expressed in E. coli, insect cells and mammalian cells. The portfolio allow optimization of process for maximum protein yield, stability and solubility. The PurKine GST-Tag Purification resin is also available as prepacked spin column and kit formats. Agarose resin and magbeads formation are both available. GST Tag Fusion - Purification and Detection Tab. 1 GST Tag Resin Supplier Elution volume (ml) Elution concentration (mg/ml) Maximum binding capacity (mg GST fusion protein/ml) Tab. 1. PurKine GST-Tag Glutathione Resin 4FF featured higher elution rate and taget protein binding capacity than supplier G. Fig. 4. GST recombinant proteins were Abbkine purified with PurKine GST-Tag Glutathione Resin 4FF (Lane B) and counterpart product Supplier G from supplier G (Lane C). A B C Fig. 4 PurKine GST-Tag Glutathione Resin Agarose Resin BMR ml, 10ml, 50ml, 100ml PurKine GST-Tag Glutathione Packed Column Packed Column BMC ml 5, 3ml 5 PurKine GST-Tag Purification Kit (Glutathione) Kit (Agarose Resin) KTP ml 5, 3ml 5 PurKine GST-Tag Glutathione Resin 4FF Agarose Resin BMR ml, 10ml, 50ml, 100ml PurKine GST-Tag Glutathione Packed Column 4FF Packed Column BMC ml 5, 3ml 5 PurKine GST-Tag Glutathione Magbeads Agarose Magbeads BMM ml, 5ml, 10ml GST-Tag Protein Detection Antibodies Specific detection of GST Tag fusion proteins with GST tag monoclonal and polyclonal antibodies Anti-GST tag antibodies are widely used to monitor the progress of purification by western blotting. Antibodies are also handy in order to ensure no proteolytic degradation of the recombinant protein is taking place. Anti-GST Tag Mouse Monoclonal Antibody (2A8) WB A μl, 200μl, 1ml, 10ml GST-Tag Polyclonal Antibody WB ABP μl, 100μl, 200μl, 10ml Plant GST Rabbbit Polyclonal Antibody WB ABP μl, 100μl, 200μl, 10ml 3

6 MBP Tag Fusion - Purification and Detection MBP Tag Fusions - Purification and Detection Maltose-Binding Protein (MBP) is a native E. coli protein with 370 amino acid residues. As a fusion tag, MBP is useful for recombinant protein purification by affinity chromatography. Generally, recombinant proteins tagged with MBP can alleviate toxicity and improve expression level and protein solubility. MBP tagging may produce a higher percentage of recombinant protein than that the polyhistidine tag does. However, the disadvantage of MBP is the size and immunogenicity of the affinity tag, which complicates any downstream application. MBP-Tag Protein Purification Tools Effetive purification of MBP fusion proteins with Dextrin coupled agarose resins or magbeads The elution of the MBP-fused proteins is at neutral ph using mild maltose-containing buffer conditions. MBP tag is effective when placed on the N-terminal or C-terminal end of target proteins. Dextrin beads is a chromatography medium for purifying proteins fused to maltose binding protein (MBP-tagged protein). The small, evenly sized dextrin beads ensure that MBP-tagged proteins elute in narrow peaks, thus minimizing the need for further concentration steps. PurKine MBP-Tag Purification resin is also available as prepacked spin column and kit formats. Agarose resin and magbeads formation are both available. PurKine MBP-Tag Dextrin Resin 6FF Agarose Resin BMR ml, 10ml, 50ml, 100ml PurKine MBP-Tag Dextrin Packed Column 6FF Packed Column BMC ml 5, 3ml 5 PurKine MBP-Tag Protein Purification Kit (Dextrin) Kit (Agarose Resin) KTP ml 5, 3ml 5 PurKine MBP-Tag Dextrin Magbeads Agarose Magbeads BMM ml, 5ml, 10ml Fig. 5 Fig. 5. PurKine MBP-Tag Dextrin resin (Lane C) and MBP Tag Amylose resin from supplier N (Lane D) were both applied for MBP tag recombinant protein purification with the same condition. PurKine MBP-Tag Dextrin performs better both in target protein productivity and purity. Fig. 6 Fig. 6. Western blot analysis of 0.5ug MBP fusion protein with Anti-MBP Tag Mouse Monoclonal Antibody (9Y5) in 1:2,000 (Lane A), 1:3,000 (Lane B) and 1:5,000 (Lane C) dilutions. A B C D A B C MBP-Tag Protein Detection Antibodies Specific detection of MBP Tag fusion proteins with MBP tag monoclonal and polyclonal antibodies Anti-MBP tag antibodies are widely used to monitor the progress of purification by western blotting, and other assay methods involving MBP-tagged proteins. Anti-MBP Tag Mouse Monoclonal Antibody (9Y5) WB A μl, 200μl, 1ml, 10ml MBP-Tag Polyclonal Antibody WB ABP μl, 100μl, 200μl, 10ml 4

7 DDDDK Tag Fusions - Purification and Detection DDDDK-tag (Flag tag) is a polypeptide protein tag that can be added to a protein using recombinant DNA technology, having the sequence motif DYKDDDDK, which allows elution under non-denaturing conditions. It has been used for studying proteins in living cells and for protein purification by affinity chromatography. It can also be used in the isolation of protein complexes with multiple subunits, because its mild purification procedure tends not to disrupt such complexes. DDDDK Tag Protein Purification Tools Specific purification of DDDDK tag fusion proteins with anti-ddddk antibody coupled resin Anti-DDDDK Resin 4FF resin, coupled with mouse monoclonal antibody against DYKDDDDK tag, has been designed for affinity purification of Flag fusion proteins. DDDDK Tag Fusion - Purification and Detection PurKine Anti-DDDDK Tag Resin 4FF Agarose Resin BMR ml, 5ml, 50ml DDDDK Tag Protein Detection Antibodies Specific detection of DDDDK Tag fusion proteins with WB, IF and IP immunoassays The DDDDK tag is likely to be located on the surface of a fusion protein because of its hydrophilic nature and therefore is more likely to be accessible to antibodies. DDDDK tag binds to several specific anti-ddddk antibodies with different recognition and binding characteristics. The recombinant DDDDK fusion protein can be recognized by this high-affinity 1B10 antibody. Anti-DDDDK Tag Mouse Monoclonal Antibody (1B10) WB, IF, IP A μl, 200μl, 1ml, 10ml Anti-DDDDK Tag Rabbit Polyclonal Antibody WB A μl, 200μl, 1ml, 10ml HRP Conjugated Anti-DDDDK Tag Mouse Mab (1B10) WB A μl, 200μl, 1ml, 10ml Fig. 7 Fig. 7. IP (1:200) - WB (1:5,000) analysis of Flag fusion protein expression in 293 cells. Fig. 8 A B C D E Untransfected 293 cell lysate (lane A), transfected 293 cell lysate with Flag-tag protein (lane B), IP transfected 293 with normal Mouse IgG and Protein G agarose (lane C), IP transfected 293 with Anti Flag tag mab and Protein G agarose (lane D), and IP transfected 293 with only Protein G agarose (lane E). Fig. 8. IF staining (1:2,000) of DDDDK tag fusion protein in 293 cells with Anti-DDDDK Tag Mouse Monoclonal Antibody (1B10) and IFKine Red Donkey anti-mouse secondary antibody. Blue color shows counterstained part with DAPI. DDDDK Tag Blocking Peptide High quality blocking peptide for Anti-DDDDK tag antibody DDDDK Tag Peptide WB, BL PRQ mg, 20mg, 100mg 5

8 Biotin Tag Fusions - Purification, Detection Biotin Tag Fusions - Purification and Detection Comparing with other tags, biotin is comparatively smaller than globular proteins, which minimizes any significant interference in many proteins and allows multiple biotin molecules to be conjugated to a single protein for maximum detection by streptavidin. Also, biotin has a valeric acid side chain that is easily derivatized and conjugated to reactive moieties and chemical structures without affecting its avidin-binding function. Biotin-Tag Protein Purification Tools Effetive purification of biotinylated proteins using streptavidin resin and magbeads PurKine Biotin-Tag purification system is based on innovative high-capacity matrix for convenient single-step purifications of biotin and biotinylated substances. Biotinylated antibodies, proteins, peptides, nucleic acids and other molecules or interaction complexes can be purified from samples using this streptavidin resin. Agarose resin and magbeads formation are both available. PurKine Biotin-Tag Streptavidin Resin 6FF Agarose Resin BMR ml, 10ml, 50ml, 100ml PurKine Biotin-Tag Streptavidin Packed Column 6FF Packed Column BMC ml 5, 3ml 5 PurKine Biotin-Tag Protein Purification Kit (Streptavidin) Kit (Agarose Resin) KTP ml 5, 3ml 5 PurKine Biotin-Tag Streptavidin Magbeads Agarose Magbeads BMM ml, 5ml, 10ml Biotin-Tag Protein Detection Tools Specific detection of biotinylated proteins with HRP conjugated streptavidin or anti-biotin antibodies HRP, Streptavidin WB, IHC, ELISA A ul, 500ul HA Tag Fusions - Purification, Detection and Blocking HA tag comes from human influenza hemagglutinin (HA) corresponding to amino acids and is a strong immunoreactive epitope making it popular to isolate, purify, detect, and track the protein of interest. The recombinant HA-tagged proteins can be separated by highly specific anti- HA monoclonal antibody that is covalently immobilized on resin. PurKine Anti-HA Tag Resin 4FF Agarose Resin BMR ml, 5ml, 50ml Anti-HA Tag Mouse Monoclonal Antibody (4F6) WB, IF, IP A μl, 200μl, 1ml, 10ml Anti-HA Tag Rabbit Polyclonal Antibody WB, IP ABP μl, 100μl, 200μl, 10ml HRP Conjugated Anti-HA Tag Mouse Mab (4F6) WB A μl, 100μl, 200μl, 10ml HA Tag Peptide WB, BL PRQ mg, 20mg, 100mg 6

9 Protein A Tag Fusions - Purification Protein A tags are used in tandem affinity purification (TAP), a technique for studying protein-protein interactions. TAP differs from standard affinity purification by using two successive affinity chromatography steps to enhance the specificity of the purification procedure and reduce the number of false positives. The matrix of PurKine Protein A-Tag IgG Resin 4FF is covalently coupled to human IgG. This resin allows high flow rates, for rapid and convenient purification of protein A fusion protein conjugates. Protein A-Tag Protein Purification Tools Effetive purification of protein A fusion proteins using this human IgG resin Protein A Tag Fusion - Purification PurKine Protein A-Tag IgG Resin 4FF Agarose Resin BMR ml, 10ml, 50ml, 100ml Myc Tag Fusions - Detection and Blocking Myc tag originates from the c-myc gene product with peptide sequence EQKLISEEDL. The recombinant Myc fusion protein can be recognized by a well-known high-affinity 2D5 antibody, which can specifically recognizes native and denatured forms of Myc fusion proteins, with which can meet your any requirements for Western Blot, Immunofluorescence and Immunoprecipitation assays. Under native conditions, the elution of Myc-tag proteins can be achieved by the addition of the Myc tag peptide which competes with the recombinant proteins. Anti-Myc Tag Mouse Monoclonal Antibody (2D5) WB, IF, IP A μl, 200μl, 1ml, 10ml Anti-Myc Tag Rabbit Polyclonal Antibody WB A μl, 200μl, 1ml, 10ml HRP Conjugated Anti-Myc Tag Mouse Mab (2D5) WB A μl, 200μl, 1ml, 10ml Myc Tag Peptide WB, BL PRQ mg, 20mg, 100mg VSV-G Tag Fusions - Detection and Blocking VSV-G, a vesicular stomatitis virus G (VSV-G) protein fragment, which is commonly used in biomedical research to pseudotype retroviral and lentiviral vectors. The VSV-G epitope tag is commonly engineered onto the N- or C- terminus of a protein of interest so that the tagged protein can be analyzed and visualized using immunochemical methods. Anti-VSV-G-Tag Mouse Monoclonal Antibody (14D2) WB, IF, IP A μl, 200μl, 1ml, 10ml VSV-G Tag Peptide WB, BL PRQ mg, 20mg, 100mg 7

10 Other Tag Fusions - Detection and Blocking Other Tag Fusions - Detection and Blocking Other Tag Protein Detection Antibodies Full portfolio of tag specific monoclonal and polyclonal antibodies The use of epitope tags eliminates the additional step of using a different antibody for each newly purified protein. Besides the most commonly used epitope tags include His, HA, Myc, GST and DDDDK tag, Abbkine offers highly specific antibodies to all of these epitope tags. Our tagged antibodies are manufactured under extensive quality control to ensure optimal sensitivity and consistent high performance, which can serve as a universal detection method in applications such as western blot, immunofluorescence, and immunoprecipitation. Our customer service team also provide you customerized tag fusion antibodies production, tag peptides synthesis and modification services per your request. Anti-mCherry Tag Mouse Monoclonal Antibody (9D3) WB, IF A μl, 200μl, 1ml, 10ml Anti-KT3 Tag Mouse Monoclonal Antibody (14D8) WB A μl, 200μl, 1ml, 10ml Anti-RFP Tag Mouse Monoclonal Antibody (9D1) WB A μl, 200μl, 1ml, 10ml Anti-V5 Tag Mouse Monoclonal Antibody (11D5) WB, IF, IP A μl, 200μl, 1ml, 10ml Anti-CBP Tag Monoclonal Antibody (12H5) WB A μl, 200μl, 1ml, 10ml Anti-TAP Tag Monoclonal Antibody (4H2) WB A μl, 200μl, 1ml, 10ml Anti-Avi-Tag Monoclonal Antibody (5G11) WB A μl, 200μl, 1ml, 10ml Anti-SRT-Tag Monoclonal Antibody (11G3) WB A μl, 200μl, 1ml, 10ml Anti-mStrawberry Monoclonal Antibody (4C9) WB A μl, 200μl, 1ml, 10ml Anti-EYFP Monoclonal Antibody (10T3) WB A μl, 200μl, 1ml, 10ml Anti-mOrange Monoclonal Antibody (9A10) WB A μl, 200μl, 1ml, 10ml Anti-AmCyan Monoclonal Antibody(8T2) WB A μl, 200μl, 1ml, 10ml Anti-ECFP-Tag Monoclonal Antibody(6B11) WB A μl, 200μl, 1ml, 10ml Anti-EBFP Monoclonal Antibody(8B5) WB A μl, 200μl, 1ml, 10ml Anti-Nano-Tag9 Monoclonal Antibody(11T3) WB A μl, 200μl, 1ml, 10ml Other Tag Blocking Peptides High quality blocking peptide for tag antibody activities with affordable price Synthetic epitope peptides is used for tag antibody production after KPL conjugation, or blocking their counterpart antibody activities. Some researchers also use tag peptide to competes with its counterpart recombinant proteins in their cellular analysis. T7 Tag Peptide WB, BL PRQ mg, 20mg, 100mg S Tag Peptide WB, BL PRQ mg, 20mg, 100mg V5 Tag Peptide WB, BL PRQ mg, 20mg, 100mg 8

11 Protein A Resin and Magbeads Protein A is a 42 kda surface protein originally found in the cell wall of the bacteria Staphylococcus aureus, which is composed of five homologous immunoglobulin-binding domains. Protein A binds the heavy chain within the Fc region of most immunoglobulins and also within the Fab region in the case of the human VH3 family. Through these interactions in serum, where IgG molecules are bound in the wrong orientation, the bacteria disrupts opsonization and phagocytosis. Protein A for Antibody Purification Protein A Resin and Magbeads PurKine Protein A Purification system is designed for easy, one-step purification of classes, subclasses and fragments of immunoglobulins from biological fluids and from cell culture media. Recombinant protein A is produced in E. coli and functions essentially the same as native Protein A from Staphylococcus aureus. The Protein A molecule contains four high-affinity binding sites capable of interacting with the Fc region from IgG of several species. Protein A is generally preferred for affinity purification of rabbit, guinea pig, dog and cat IgG. The portfolio allow optimization of process for maximum protein yield, stability and solubility. The PurKine Protein A Purification Resin is also available as prepacked spin column and kit formats. Agarose resin and magbeads formation are both available. Tab. 2 Protein A Resin Supplier Elution volume (ml) Elution concentration (mg/ml) Maximum binding capacity (mg higg/ml) Tab. 2. PurKine Protein Resin 4FF featured higher elution rate and taget protein binding capacity than supplier G. Fig. 9. Human IgG were purified with Abbkine PurKine Protein A Resin 4FF (Lane B) and counterpart product from supplier G (Lane Supplier G C). A B C Fig. 9 PurKine Protein A Resin Agarose Resin BMR ml, 10ml, 50ml, 100ml PurKine Protein A Packed Column Packed Column BMC ml 5, 3ml 5 PurKine Antibody Purification Protein A Kit Kit (Agarose Resin) KTP ml 5, 3ml 5 PurKine Protein A Resin 4FF Agarose Resin BMR ml, 10ml, 50ml, 100ml PurKine Protein A Magbeads Agarose Magbeads BMM ml, 5ml, 10ml Protein At (Alkaline tolerate) for Antibody Purification PurKine Protein AT Purification system is effective for affinity purification of monoclonal antibodies at process scale. Protein At is an alkali-resistant protein A (Alkaline tolerate) based on the native protein A though biotechnology mutation. This ligand provides greater stability than conventional protein A, and cleaning can be performed with cost-effective reagents such as sodium hydroxide eliminating the need for expensive and hazardous cleaning agents. The PurKine Protein AT Purification Resin is also available as prepacked spin column and kit formats. PurKine Protein AT Resin 4FF Agarose Resin BMR ml, 10ml, 50ml, 100ml PurKine Protein AT Packed Column 4FF Packed Column BMC ml 5, 3ml 5 9

12 Protein G Resin and Magbeads Protein G Resin and Magbeads Protein G for Antibody Purification Protein G is an immunoglobulin-binding protein expressed in group C and G Streptococcal bacteria much like Protein A but with differing binding specificities. It is a 65-kDa (G148 protein G) and a 58 kda (C40 protein G) cell surface protein that has found application in purifying antibodies through its binding to the Fab and Fc region. The native molecule also binds albumin, but because serum albumin is a major contaminant of antibody sources, the albumin binding site has been removed from recombinant forms of Protein G. PurKine Protein G Purification system is effective for affinity purification of IgG from serum and other fluids of many mammalian species. Protein G can be used for purification of mammalian monoclonal and polyclonal IgGs that do not bind well to protein A. Protein G has greater affinity than protein A for most mammalian IgGs, especially for certain subclasses including human IgG3, mouse IgG1 and rat IgG2a. Unlike protein A, protein G does not bind to human IgM, IgD and IgA. The PurKine Protein G Purification Resin is also available as prepacked spin column and kit formats. Agarose resin and magbeads formation are both available. PurKine Protein G Resin Agarose Resin BMR ml, 10ml, 50ml, 100ml PurKine Protein G Packed Column Packed Column BMC ml 5, 3ml 5 PurKine Antibody Purification Protein G Kit Kit (Agarose Resin) KTP ml 5, 3ml 5 PurKine Protein G Resin 4FF Agarose Resin BMR ml, 10ml, 50ml, 100ml PurKine Protein G Magbeads Agarose Magbeads BMM ml, 5ml, 10ml Protein L Resin Protein L is an immunoglobulin-binding protein that was originally derived from the bacteria Peptostreptococcus magnus. Unlike Protein A and Protein G, Protein L binds antibodies through light chain interactions. Since no part of the heavy chain is involved in the binding interaction, Protein L binds a wider range of antibody classes than Protein A or G. Protein L binds to representatives of all antibody classes, including IgG, IgM, IgA, IgE and IgD. However, Protein L binding is restricted to those antibodies that contain kappa light chains. Protein L for Antibody Purification PurKine Protein L Purification system is effective for affinity purification of mammalian IgG that contain specific kappa light chains from serum, ascites fluid, cell culture supernantant and other antibody samples. Recombinant Protein L is chemically modified with a proprietary method to minimize nonspecific binding of proteins, which is extremely useful for purification of VLk-containing monoclonal antibodies from culture supernatant because it does not bind bovine immunoglobilins, which are present in the media serum supplement. PurKine Protein L Resin Agarose Resin BMR ml, 10ml, 50ml, 100ml PurKine Protein L Packed Column Packed Column BMC ml 5, 3ml 5 PurKine Protein L Resin 4FF Agarose Resin BMR ml, 10ml, 50ml, 100ml PurKine Protein L Packed Column 4FF Packed Column BMC ml 5, 3ml 5 10

13 Protein A/G Resin and Magbeads Protein A/G is a recombinant fusion protein that combines IgG binding domains of both Protein A and Protein G. It binds to all subclasses of human IgG, making it useful for purifying polyclonal or monoclonal IgG antibodies whose subclasses have not been determined. In addition, it binds to IgA, IgE, IgM and IgD. Protein A/G also binds to all subclasses of mouse IgG but does not bind mouse IgA, IgM or serum albumin. This allows Protein A/G to be used for purification and detection of mouse monoclonal IgG antibodies, without interference from IgA, IgM and serum albumin. Protein A/G Resin and Magbeads Protein A/G for Antibody Purification PurKine Protein A/G Purification system is effective for affinity purification of IgG from serum, ascites fluid, cell culture supernantant and other antibody samples. Recombinant Protein A/G is chemically modified with a proprietary method to minimize nonspecific binding of proteins. Also, Protein A/G binding to immunoglobulins is not as ph-dependant as Protein A. Protein A/G binds to all human IgG subclasses and binds well to all mouse IgG subclasses but does not bind mouse IgA, IgM or serum albumin. The PurKine Protein A/G Purification Resin is also available as prepacked spin column and kit formats. Agarose resin and magbeads formation are both available. PurKine Protein A/G Magbeads are typically used for small-scale isolating antibodies from serum, cell culture supernatant or ascites and for immunoprecipitation and co-immunoprecipitation of antigens from cell or tissue extracts. PurKine Protein A/G Resin 4FF Agarose Resin BMR ml, 10ml, 50ml, 100ml PurKine Antibody Purification Protein A/G Kit Kit (Agarose Resin) KTP ml 5, 3ml 5 PurKine Protein A/G Magbeads Agarose Magbeads BMM ml, 5ml, 10ml Antibody Immunoaffinity Purification Resin Antigen-specific antibody purification is the immunoaffinity chromatography method for most researchers when antibodies, especially polyclonal antibodies have been generated against peptides. Firstly, purified ligand molecules are immobilized to a solid support, then a complex mixture containing the target molecules is added to the solid support. After washing away nonbound components of the complex mixture, the captured target molecules are released and recovered. This process enables maximum isolation and enrichment of only the most specific antibodies generated to the antigen while removing other unwanted pre-immune antibodies. PurKine Protein SulfyBind Resin is a simple and efficient affinity chromatography medium, which allows covalent immobilization of sulfhydryl containing peptides, protein and other ligands to the medium agarose support for use in affinity purification procedures. While PurKine Protein AminoBind Resin 4FF containing N-hydroxysuccinimide (NHS) functional groups, forms a chemically stableamide bone with ligands containing primary amino groups. Thus can be used to prepare affinity purification procedures which can isolate specific substances from complex mixtures, often achieving very high purity in a single step. PurKine Protein SulfyBind Resin Agarose Resin BMR ml, 10ml, 50ml, 100ml PurKine Protein AminoBind Resin 4FF Agarose Resin BMR ml, 10ml, 50ml, 100ml 11

14 IPKine Specific Chain Secondary Antibodies IPKine Specific Chain Secondary Antibodies When labeled secondary antibodies specific for both heavy and light chains of IgG are used to detect protein bands on Western blots following Immunoprecipitation. The secondary antibody will recognize the primary antibody used in WB as well as the denatured/reduced primary antibody released during the IP procedure. It directly result in two bands: the heavy chain (50kDa) and the light chain (25kDa). These bands usually obscure detection of any protein of interest with a molecular weight near 50 kda or 25 kda. Abbkine IPKine sencondary antibodies solve these problems with less background noise, and enhanced accuracy. IPKine light chain specific (LCS) and heavy chain specific (HCS) antibodies will eliminate heavy and light chain interference, respectively, while serum proteins absorbed minimize cross-reactivity with immunoglobulins from other species which may be present on blots. IPKine HRP Goat Anti-Rabbit IgG HCS WB, IP A μl, 500μl IPKine HRP Goat Anti-Mouse IgG HCS WB, IP A μl, 500μl IPKine HRP Mouse Anti-Rabbit IgG LCS WB, IP A μl, 500μl IPKine HRP Goat Anti-Mouse IgG LCS WB, IP A μl, 500μl IFKine Secondary Antibodies Immunofluorescence (IF) microscopy is a particularly robust and broadly applicable method generally used by researchers to assess both the localization and endogenous expression levels of proteins of interest. In multiple labeling IF assay, researchers commonly use primary antibodies raised in different species, and pre-adsorbed secondary antibodies conjugated to different fluorophores while from the same host species to minimize species cross-reactivity. The brightest fluorophores will help for low abundant proteins and vice versa to maximize sensitivity. IFKine is series of unique fluorence staining secondary antibodies with improved brightness, photostability and less nonspecific hybridization and background. The latest generation of IFKine fluorescent dyes ensure the best fluorescent performance, while it s donkey host and other species of serum/igg absorbed make IFKine secondary antibodies the ideal for use in fluorence staining, especially in fluorence multiple labeling. IFKine Green Donkey Anti-Mouse IgG IF, FC A ul, 500ul IFKine Green Donkey Anti-Rabbit IgG IF, FC A ul, 500ul IFKine Green Donkey Anti-Goat IgG IF, FC A ul, 500ul IFKine Orange Donkey Anti-Mouse IgG IF, FC A ul, 500ul IFKine Red Donkey Anti-Mouse IgG IF, FC A ul, 500ul IFKine Red Donkey Anti-Rabbit IgG IF, FC A ul, 500ul 12

15 Part of Abbkine Product Publications Abbkine products have been widely cited and recommended by hundreds and thousands of world s life science research journals. Below are part of publications list. If you have published articles using Abbkine s products, kindly send to marketing@abbkine. com, or submit your publications online here. We ll reward you gift for your continue research. Xu H J, Xue J, Lu B, et al. Two insulin receptors determine alternative wing morphs in planthoppers. Nature, 2015, 519(7544): IF: Huang, Hai-Jian, et al. Rice ragged stunt virus-induced apoptosis affects virus transmission from its insect vector, the brown planthopper to the rice plant. Scientific reports 5 (2015). IF: Yu L, Liang H, Lu Z, et al. Membrane receptor-dependent Notch1/Hes1 activation by melatonin protects against myocardial ischemia reperfusion injury: in vivo and in vitro studies. Journal of pineal research, 2015, 59(4): IF: Chen G, Tian F, Li C, et al. In vivo real-time visualization of mesenchymal stem cells tropism for cutaneous regeneration using NIR-II fluorescence imaging. Biomaterials, 2015, 53: IF: Li C, Cao L, Zhang Y, et al. Preoperative Detection and Intraoperative Visualization of Brain Tumors for More Precise Surgery: A New Dual Modality MRI and NIR Nanoprobe. Small, 2015, 11(35): IF: Liu S L, Wu Q M, Zhang L J, et al. Three-Dimensional Tracking of Rab5- and Rab7- Associated Infection Process of Infl uenza Virus. Small, 2014, 10(22): IF: Long Y M, Zhao X C, Clermont A C, et al. Negatively charged silver nanoparticles cause retinal vascular permeability by activating plasma contact system and disrupting adherens junction. Nanotoxicology, 2016, 10(4): IF: Xue X, Wang Q, Qu Y, et al. Development of the photosynthetic apparatus of Cunninghamia lanceolata in light and darkness. New Phytologist, IF: Li Y, Zhang L, Li D, et al. The Arabidopsis F-box E3 ligase RIFP1 plays a negative role in abscisic acid signalling by facilitating ABA receptor RCAR3 degradation. Plant, cell & environment, 2016, 39(3): IF: Niu, Xuan, et al. P2Y12 Promotes Migration of Vascular Smooth Muscle Cells Through Cofilin Dephosphorylation During Atherogenesis. Arteriosclerosis, Thrombosis, and Vascular Biology (2017): ATVBAHA-116. IF: Zheng, Xiaoyao, et al. A hybrid sirna delivery complex for enhanced brain penetration and precise amyloid plaque targeting in Alzheimer s disease mice. Acta Biomaterialia (2016). IF: Ji Y X, Zhang P, Zhang X J, et al. The ubiquitin E3 ligase TRAF6 exacerbates pathological cardiac hypertrophy via TAK1-dependent signalling. Nature communications, 2016, 7. IF: Zheng, Peng, et al. Quantitative Proteomics Analysis Reveals Novel Insights into Mechanisms of Action of Long Noncoding RNA Hox Transcript Antisense Intergenic RNA (HOTAIR) in HeLa Cells. Molecular & Cellular Proteomics 14.6 (2015): IF: Yang L, Tang H, Kong Y, et al. LGR5 Promotes Breast Cancer Progression and Maintains Stem-Like Cells Through Activation of Wnt/β-Catenin Signaling. Stem Cells, 2015, 33(10): IF: Wan, Quanyuan, et al. MDA5 Induces a Stronger Interferon Response than RIG-I to GCRV Infection through a Mechanism Involving the Phosphorylation and Dimerization of IRF3 and IRF7 in CIK Cells. Frontiers in Immunology 8 (2017): 189. IF: Li H T, Zhao X Z, Zhang X R, et al. Exendin-4 Enhances Motor Function Recovery via Promotion of Autophagy and Inhibition of Neuronal Apoptosis After Spinal Cord Injury in Rats. Molecular neurobiology, 2015: IF: Fang Z, He Q W, Li Q, et al. MicroRNA-150 regulates blood brain barrier permeability via Tie-2 after permanent middle cerebral artery occlusion in rats. The FASEB Journal, 2016, 30(6): IF: Zheng T, Yang X, Wu D, et al. Salidroside ameliorates insulin resistance through activation of a mitochondria-associated AMPK/PI3K/Akt/GSK3β pathway. British journal of pharmacology, 2015, 172(13): IF: Lv, Meinan, et al. Characterization of a C3 Deoxygenation Pathway Reveals a Key Branch Point in Aminoglycoside Biosynthesis.Journal of the American Chemical Society (2016): IF: Liu, L, et al. Baclofen mediates neuroprotection on hippocampal CA1 pyramidal cells through the regulation of autophagy under chronic cerebral hypoperfusion. Scientific Reports 5(2015): IF: Yang, Dongxue, et al. NPAS3 regulates transcription and expression of VGF: implications for neurogenesis and psychiatric disorders. Frontiers in Molecular Neuroscience 9 (2016). IF:

16 Abbkine Scientific Co., Ltd. was founded by a number of scientists and marketing experts in the field of life science in California, USA in With growing demands from Asia Pacific, it move its headquarters to China. Combining cutting edge technology from United States with China s manufacturing engineering and cost advantages, we aim to provide innovative, high quality assay kits, recombinant proteins, antibodies and other research tools to accelerate life science fundamental research, drug discovery, etc. Contact us Tel: Fax: service@abbkine.com Or your local distributors Our mission is to help make research possible by supplying scientists worldwide with the basic research tools necessary for advancing human and animal health. We would achieve these goals through a well-defined core strategy and a company culture that fosters integrity, openness, and collaboration, and rewards customer focus, innovation and ownership, thus offer global life science researchers with the highest quality products with an affordable pricing policy.