Catalog # HM0501

Anti-His (C-terminal) Tag Antibody

Mouse Monoclonal

Application / Dilution
ELISA (Cap)1:200
ICC1:100
IHC1:100
IP1:100
WB1:5000

Size 100 μl

Isotype IgG1

$275


DATASHEET  

Epitope tagging (e.g. His-tag) is a technique in which a known epitope is fused to a recombinant protein using genetic engineering. By choosing a particular epitope and recombinant protein combination, epitope tagging makes it possible to detect proteins for which no antibody is available. The 6x His tag is a synthetic oligo peptide consisting of 6 consecutive histidine residues (HHHHHH). A variety of plasmids contain DNA that encodes an amino- or carboxy-terminal tag consisting of six histidine (6xHis) residues followed by an extended multiple cloning site. These plasmids facilitate expression of His-tagged recombinant proteins that can be isolated or purified by immobilized metal affinity chromatography. Anti-His Tag antibodies can be used to detect recombinant proteins with the His-Tag in many different immunoassays, including western blot, immunoprecipitation, immunocytochemistry, immunohistochemistry, flow cytometry, ELISA, and chromatin immunoprecipitation assays.

References

Kroll, D.J. et al. (1993) DNA Cell Biol. 12:441.

Western blot of human recombinant DDR1 protein with C-terminal His Tag. The blot was probed with mouse monoclonal anti-His (Cterminal) Tag (HM0501) at 1:2000 (lane 1), 1:4000 (lane 2), and 1:8000 (lane 3).

Immunocytochemical labeling of Axl recombinant protein with C-terminal His Tag bound to THP1 aldehyde fixed cells. The cells were labeled with mouse monoclonal anti-His (C-terminal) Tag antibody (HM0501). The antibody was detected using goat anti-mouse DyLight® 594.



Representative Standard Curve using mouse monoclonal anti-His (C-terminal) Tag antibody (HM0501) for ELISA capture of human recombinant CD46 extracellular region with a C-terminal His-tag. Captured protein was detected using anti-CD46 antibody (CM0371) antibody followed by appropriate secondary antibody HRP conjugate.

Clone (M050) was generated from a recombinant protein containing a C-terminal His-tag.

Mouse monoclonal antibody purified with protein G chromatography is supplied in 100 µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.

Clone M050 detects recombinant proteins with C-terminal His-tag epitope. The antibody binds 6 and 10 His-tag epitopes on the C-terminal side of a protein, but does not bind N-terminal His-tag epitopes. The antibody works for western blot, immunoprecipitation, immunocytochemistry, flow cytometry, and ELISA capture.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

This kit contains: