THE® Anti-His Monoclonal Antibody PRODUCT INFORMATION Description Monoclonal antibodies specific to polyhistidine can greatly improve the effectiveness of several different kinds of immunoassays, helping researchers identify, detect, and purify polyhistidine fusion proteins in bacteria, insect cells, and mammalian cells. However, since 6xHis-tag is poorly immunogenic, it needs to be conjugated to KLH or some other carrier as an immunogen. After hundreds of selection cycles, researchers at GenScript successfully isolated an antibody against His-tag. THE® Anti-His mAb (subtype IgG1) has very high affinity. Tests performed at GenScript show that the antibody can also recognize 4xHis- and 5xHis-tags. This means that even if the 6xHis-tag is only partially exposed, it will still be recognized and bound by this antibody.THE ® Anti-His mAb is produced from mice ascites and purified by protein G affinity column. This antibody recognizes native as well as denatured forms of synthetic polyhistidine and polyhistidine-tagged fusion proteins. The product reacts with fusion proteins expressed in bacteria, insect cells, and mammalian cells. THE ® Anti-His mAb recognizes polyhistidine tags placed at N-terminal, C-terminal, and internal regions of fusion proteins. THE® Anti-His mAb can be used in Western blot analyses, Dot blot analyses, ELISA, immunofluorescent staining, and flow cytometry of cultured cells. Cat. No.: A00186 Host: Mouse Size: 100 ug Immunogen: Synthetic peptide containing six histidine residues (6xHis) coupled to KLH Antigen Synonyms: Histidine Ig Subclass: Mouse IgG1 Clone: A00186.01 Purification: Protein G affinity column Conjugation: Unconjugated Specificity The antibody recognizes C-terminal, N-terminal, and internal tagged fusion proteins. Fusion Partner Spleen cells were fused with SP2/0-Ag14 mouse myeloma cells. Concentration 100 μg, lyophilized with PBS, pH 7.4, containing 0.02% sodium azide. Reconstitution Reconstitute lyophilized THE® His mAb (100 μg) with 100 μl of deionized water to a final antibody concentration of 1 mg/ml. Storage THE® Anti-His mAb should be stored lyophilized until use. It remains stable in lyophilized form for at least two years if stored between 2°C and 8°C. The reconstituted antibody can be stored for up to three months between 2°C and 8°C or for up to two years at or below -20°C. Avoid repeated freezing and thawing cycles. Applications Working concentrations for specific applications should be determined by the investigator. Appropriate concentrations will be affected by several factors, including secondary antibody affinity, antigen concentration, sensitivity of detection method, temperature, and length of incubations, etc. The suitability of this antibody for applications other than those listed below has not been determined. The following concentration ranges are recommended starting points for this product. ELISA: 0.05µg/ml–0.2µg/ml Western blot: 0.1µg/ml–0.2µg/ml Immunoprecipitation (IP): 1µg/ml Immunofluorescent staining: 1µg/ml FAC assay/Flow cytometry: 1µg/ml ** For non-clinical research use only. ** Tel: 732-885-9188 Fax: 732-210-0262 E-Mail: info@genscript.com www.genscript.com Other applications: user-optimized Note Small volumes of antibody will occasionally become entrapped in the seal of the product vial during shipment and storage. Before use, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container's cap. Example ** For non-clinical research use only. ** Tel: 732-885-9188 Fax: 732-210-0262 E-Mail: info@genscript.com www.genscript.com