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His-Tag (27E8) Mouse mAb #2366
| Applications | Reactivity | Sensitivity | Isotype |
|---|---|---|---|
| W IP | All | Transfected Only | Mouse IgG1 |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
Reactivity Key: All=All species expected
Specificity / Sensitivity
His-Tag (27E8) Mouse mAb only recognizes His-tag. It is expected that this antibody will react with proteins tagged with His from all species.
Source / Purification
Monoclonal antibody is produced by immunizing mice with a 6xHis synthetic peptide (KLH-coupled).
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 usefull 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.
- di Guan, C. et al. (1988) Gene 67, 21-30.
Application References
- Rishi, A. K. et al. (2003) Identification and characterization of a cell cycle and apoptosis regulatory protein-1 as a novel mediator of apoptosis signaling by retinoid CD437. J. Biol. Chem. 278, 33422-33435. This article references the use of His-Tag (27E8) Mouse mAb in the following applications: IC-ABC Western Blotting
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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.
