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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

Specificity / Sensitivity

His-Tag Antibody detects His-tagged recombinant proteins.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a 6xHis synthetic peptide (KLH-coupled). Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

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

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

IF-IC

Immunocytochemical staining of COS cells transfected with N-terminal His-tagged protein (left) or cells transfected with control cDNA (right), using His-Tag Antibody.


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 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).

  1. Kroll, D. J. et al. (1993) DNA Cell Biol. 12, 441-453.
  2. di Guan, C. et al. (1988) Gene 67, 21-30.

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This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.

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