His Tag Antibody [HRP]

Product Discontinued

His Tag Antibody [HRP] Summary

• Immunogen: Synthetic peptide: HHHHHH conjugated to KLH.
• Specificity: Chicken Polyclonal to Polyhistidine tag
• Clonality: Polyclonal
• Host: Chicken
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • ELISA 1:5000
  • Western Blot 1:500
• Application Notes: ELISA: a 1/10000 dilution gave an O.D.= 1.0 in a 15 minute reaction against peptide conjugated to a different carrier than used for anti-peptide purification. Western Blot: use at dilution of 1/1000.

Reactivity Notes

6-His Tagged proteins.

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: 50 mM HEPES (pH 7.0), 0.1 M NaCl, and 0.2% BSA
• Preservative: 0.005% Thimerosal
• Concentration: 0.5 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for His Tag Antibody [HRP]

• 6 His epitope tag
• 6-His Tag
• 6X His Tag
• 6X His
• 6x-His Tag
• 6X-His
• H
• Hexa His tag
• HHHHHH epitope tag
• HHHHHH tag
• His Tag
• HIS
• His6-Tag
• polyhistidine tag
• polyHistidine
• Poly-histidine

Background

A his-tag (also known as histidine tag or polyhistidine tag) is a common epitope tag that typically consists of at least 6 histidine residues fused to either the carboxyl (C-) or amino (N-) terminus of a targeted recombinant protein to facilitate its purification and detection (1). The most common his-tag is the hexahistidine (His6/6-His) tag which has a theoretical molecular weight of 0.8kda (1). The histidine residues readily interact with transition metal ions such as Co2+, Ni2+, Cu2+, and Zn2+, making immobilized metal-affinity chromatography (IMAC) the preferred technique for his-tag purification (1, 2). Metal ions are immobilized and bound to by His-tags in the IMAC column via the histidine imidazole ring. The tagged protein can be eluted off the column by washing with buffers containing a low concentration of imidazole (1, 2). Due to its relatively small size, low immunogenicity, versatility under denaturing conditions, and minimal interference with the structure and function of proteins, the his-tag is one of the most widely used tags for protein purification (1-3).

References

1. Malhotra, A. (2009). Tagging for protein expression. Methods in Enzymology, Guide to Protein Purification, 2nd Edition, 463, 239-258. https://doi.org/10.1016/s0076-6879(09)63016-0

2. Terpe, K. (2003). Overview of tag protein fusions: from molecular and biochemical fundamentals to commercial systems. Applied Microbiology and Biotechnology, 60(5), 523-533. https://doi.org/10.1007/s00253-002-1158-6

3. Booth, W. T., Schlachter, C. R., Pote, S., Ussin, N., Mank, N. J., Klapper, V., ... Chruszcz, M. (2018). Impact of an N-terminal polyhistidine tag on protein thermal stability. ACS Omega, 3(1), 760-768. https://doi.org/10.1021/acsomega.7b01598

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for His Tag Antibody (NB600-1474)(1)

Publication using NB600-1474

• Propheter, DC, Hsu K-L, Mahal, LK. Recombinant lectin microarrays for glycomic analysis. Methods Mol. Biol. 723:67-77. 2011-01-01 [PMID: 21370060]