Product Pathways - Related Products
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
Protocols
Specificity / Sensitivity
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.
- 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. Applications: IC-ABC Western Blotting
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Companion Products
- 2365 His-Tag Antibody
- 2272 Myc-Tag Antibody
- 2276 Myc-Tag (9B11) Mouse mAb
- 2367 HA-Tag (6E2) Mouse mAb
- 2372 β-Gal (14B7) Mouse mAb
- 2396 MBP (8G1) Mouse mAb
- 7076 Anti-mouse IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
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.
