Product Pathways - Companion Products
His-Tag (27E8) Mouse mAb (HRP Conjugate) #9991
| Applications | Reactivity | Sensitivity | Isotype |
|---|---|---|---|
| W | Transfected Only | Mouse IgG1 |
Applications Key:
W=Western Blotting
Reactivity Key:
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
- 9991:
- Western Blotting
Specificity / Sensitivity
His-Tag (27E8) Mouse mAb (HRP Conjugate) 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 COS-7 cells, mock-transfected (-) or transfected with a construct expressing C-terminal His-tagged Tyro3 (His-Tyro3; +) using His-Tag (27E8) Mouse mAb (HRP Conjugate).
Description
This Cell Signaling Technology® antibody is conjugated to the carbohydrate groups of horseradish peroxidase (HRP) via its amine groups. The HRP conjugated antibody is expected to exhibit the same species cross-reactivity as the unconjugated His-Tag (27E8) Mouse mAb #2366.
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
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Companion Products
For Research Use Only. Not For Use In Diagnostic Procedures.
