Product Pathways - Complementary Reagents
His-Tag Antibody #2365
| Applications | Reactivity | Sensitivity | Source |
|---|---|---|---|
| W IP IF-IC | All | Transfected Only | Rabbit |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key: All=All species expected
Species cross-reactivity is determined by western blot.
Protocols
* 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 Blotting
Western blot analysis of extracts from cells expressing C-terminal His-tagged protein (lane 1) or control extract (lane 2), using His-Tag Antibody.
IP
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.
IF-IC
Confocal immunofluorescent analysis of COS cells transfected with a His-tagged protein using His-Tag Polyclonal Antibody. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Background
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.
Application References
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
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This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
