His Tag Antibody (HIS.H8) [HRP]

Western Blot: His Tag Antibody (HIS.H8) [HRP] [NBP2-31055H] - Human His-tagged recombinant proteins. WB image submitted by a verified customer review.

Western Blot: His Tag Antibody (HIS.H8) [HRP] [NBP2-31055H] - Analysis of 6x-His Epitope Tag was performed by loading various amounts of E. coli lysate containing a multi-epitope tagged protein per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with an HRP-conjugated 6x-His Epitope Tag monoclonal antibody at a dilution of 1:1000 overnight at 4C on a rocking platform and washed in TBS-0.1% Tween-20. Chemiluminescent detection was performed using SuperSignal West Pico.

• Reactivity: ET
• Applications: WB, ELISA, Flow, ICC/IF, IP, CyTOF-ready
• Clone: HIS.H8
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: HRP

His Tag Antibody (HIS.H8) [HRP] Summary

• Immunogen: This His Tag Antibody (HIS.H8) was developed against a 6x His synthetic peptide.
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • ELISA
  • Flow Cytometry
  • Immunocytochemistry/ Immunofluorescence
  • Immunoprecipitation
  • Western Blot
• Readout System:
  • NovaLume Plus Chemiluminescent Reagent LUM-555

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: PBS
• Preservative: No Preservative
• Purity: Protein A purified

Alternate Names for His Tag Antibody (HIS.H8) [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 (NBP2-31055H)(1)

Publication using NBP2-31055H

• Ajam T, De I, Petkau N Et al. Alternative catalytic residues in the active site of Esco acetyltransferases Sci Rep 2020-06-20 [PMID: 32555289]
  
  Details:
  Citation using the HRP version of this antibody.

Review for His Tag Antibody (NBP2-31055H) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Verified Customer WB Human 08/22/2019

Summary

• Application: Western Blot
• Sample Tested: His-tagged recombinant proteins
• Species: Human
• Lot: UD275259-081619-H