Target His Tag Target background Epitope tags have applications in the labeling, isolation and detection of proteins using immunoblotting, Immunoprecipitation and immunostaining techniques. Epitope tags can be used, by affinity chromatography, to separate recombinant, overexpressed protein from wild-type protein expressed by the host organism. Due its small size, the protein’s biochemical properties seem not to be affected by the tag protein. Polyhistidine-tag is also known as histidine-tag, 6xHis-tag and by the trademarked name His-tag. Immunogen The antibody was raised against HHHHHH (6x His) synthetic peptide conjugated to KLH. Specificity The antibody recognizes His-tag encoded by many commercially available vectors, regardless of the tag's location in the fusion protein sequences (i.e. reacts with N-terminal, C-terminal or internal 6x His-tags) Clone ID HIS.H8 Isotype IgG2b Preservative None Format Purified with protein G, stored in PBS pH 7.4 and lyophilized. Recommend starting dilution If reconstituted with deionized water in 100 µl: WB 1:1000-3,000, IHC 1:500-2,000. Optimal dilution has to be determined by the user. Limitations Research Use Only Storage Lyophilized antibodies can be kept at 4ºC for up to 3 months and should be kept at -20ºC for long-term storage (2 years). To avoid freeze-thaw cycles, reconstituted antibodies should be aliquoted before freezing for long-term (1 year) storage (-80ºC) or kept at 4ºC for short-term usage (2 months). For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made with the assay buffer. After the maximum long-term storage period (2 years lyophilized or 1 year reconstituted) antibodies should be tested in your assay with a standard sample to verify if you have noticed any decrease in their efficacy. To limit antibody loss or degradation, BSA (final concentration 1%) and sodium azide (final concentration 0.02%) can be added to the suggested first dilution. It is important to first verify if those preservatives are compatible with your assay.