His Tag Antibody (33D10.D2.G8) [DyLight 650]

• Reactivity: ET
• Applications: WB, DB, ELISA, IHC
• Clone: 33D10.D2.G8
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: DyLight 650

His Tag Antibody (33D10.D2.G8) [DyLight 650] Summary

• Immunogen: His Tag Antibody (33D10.D2.G8) was produced in mice by repeated immunizations with 6X His epitope tag peptide H-H-H-H-H-H conjugated to KLH using maleimide.
• Specificity: This protein-A purified antibody is directed against the 6X His motif and is useful in determining its presence in various assays. This monoclonal anti-6X His tag antibody detects over-expressed proteins containing the 6X His epitope tag. To date, this antibody has reacted with all His tagged proteins so far tested. In western blotting of bacterial extracts, the antibody does not cross-react with endogenous proteins. The antibody recognizes the His-tag (His-His-His-His-His-His) fused to either the amino- or carboxy-termini of targeted proteins in transfected or transformed cells.
• Isotype: IgG1 Kappa
• Clonality: Monoclonal
• Host: Mouse
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Dot Blot
  • ELISA
  • Immunohistochemistry
  • Western Blot
• Application Notes: Optimal dilution of this antibody should be experimentally determined.

Reactivity Notes

6x His tag specific

Packaging, Storage & Formulations

• Storage: Store at 4C in the dark.
• Buffer: 50mM Sodium Borate
• Preservative: 0.05% Sodium Azide
• Purity: Protein A purified

Notes

DyLight (R) is a trademark of Thermo Fisher Scientific Inc. and its subsidiaries.

Alternate Names for His Tag Antibody (33D10.D2.G8) [DyLight 650]

• 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.