Applications Key: W=Western Blotting IP=Immunoprecipitation Reactivity Key: All=All species expected Species cross-reactivity is determined by western blot.
His-Tag (27E8) Mouse mAb detects recombinant proteins containing the 6xHis epitope tag. The antibody recognizes the 6xHis-tag fused to either the amino or carboxy terminus of targeted proteins in transfected cells.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a 6xHis synthetic peptide.
Western Blotting
Western blot analysis of extracts from control (lane 1) or cells expressing C-terminal His-tagged protein (lane 2), using His-Tag (27E8) Mouse mAb.
IP
Immunoprecipitation of control lysate (lane 1) or C-terminal His-tagged protein (lane 2), using His-Tag (27E8) Mouse mAb, followed by Western analysis with His-Tag Polyclonal Antibody #2365.
Background
Epitope tags are useful for the labeling and detection of proteins using immunoblotting, immunoprecipitation and immunostaining techniques. Due to their small size, they are unlikely to affect the tagged protein's biochemical properties.
A variety of plasmids contain DNA that encodes an amino-terminal tag cosisting of six histidine (6xHis) residues followed by an extended multiple cloning site. The 6xHis tag on the expressed recombinant proteins allows for efficient coupling to Ni2+ affinity resins and purification by single step chromatography (1).As is the case with other protein tag systems (2), this polyhistidine tag can often be cleaved at sites recognized by proteases such as thrombin and enterokinases to isolate the protein of interest (1).
Kroll, D.J. et al. (1993) DNA Cell Biol. 12, 441-453.