His-Tag Antibody #2365
|2365S||100 µl (10 western blots)||---||In Stock||---|
|2365||carrier free and custom formulation / quantity||email request|
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Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry)
* Product-specific protocol.
Specificity / Sensitivity
His-Tag Antibody 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
Polyclonal antibodies are produced by immunizing animals with a 6xHis synthetic peptide. Antibodies are purified by protein A and peptide affinity chromatography.
Western blot analysis of extracts from cells expressing C-terminal His-tagged protein (lane 1) or control extract (lane 2), using His-Tag Antibody.
Immunoprecipitation of C-terminal His-tagged protein (lane 1) or N-terminal HA-tagged protein (lane 2), using His-Tag Antibody, then western analysis with the same antibody.
Confocal immunofluorescent analysis of COS cells transfected with a His-tagged protein using His-Tag Polyclonal Antibody. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Epitope tags are useful for the labeling and detection of proteins using immunoblotting, immunoprecipitation, and immunostaining techniques. Because of 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 consisting 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.
- di Guan, C. et al. (1988) Gene 67, 21-30.
- Dovas, A. et al. (2010) J Biol Chem 285, 23296-308. Applications: IF-IC (In Cells).
- Huang, S.H. et al. (2011) J Neurosci 31, 10602-14. Applications: Western Blotting.
- Ozawa, A. et al. (2011) Mol Endocrinol 25, 776-84. Applications: Western Blotting.
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