His Tag Antibody (HIS.H8) [HRP]

Novus Biologicals | Catalog # NBP2-31055H

Novus Biologicals
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Key Product Details

Species Reactivity

Epitope Tag

Applications

Western Blot, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, CyTOF-ready

Label

HRP

Antibody Source

Monoclonal Mouse IgG2B Clone # HIS.H8
View all His Tag Primary Antibodies »

Product Specifications

Immunogen

This His Tag Antibody (HIS.H8) was developed against a 6x His synthetic peptide.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for 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.

Applications for His Tag Antibody (HIS.H8) [HRP]

Application
Recommended Usage

CyTOF-ready

Optimal dilutions of this antibody should be experimentally determined.

ELISA

Optimal dilutions of this antibody should be experimentally determined.

Flow Cytometry

Optimal dilutions of this antibody should be experimentally determined.

Immunocytochemistry/ Immunofluorescence

Optimal dilutions of this antibody should be experimentally determined.

Immunoprecipitation

Optimal dilutions of this antibody should be experimentally determined.

Western Blot

Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 1 review rated 5 using NBP2-31055H in the following applications:

Spectra Viewer

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Spectra Viewer
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS

Preservative

No Preservative

Concentration

Concentrations vary lot to lot. See vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C. Do not freeze.

Background: His Tag

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

Long Name

Histidine Tag

Alternate Names

polyHistidine

Additional His Tag Products

Product Documents for His Tag Antibody (HIS.H8) [HRP]

Certificate of Analysis

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Product Specific Notices for His Tag Antibody (HIS.H8) [HRP]

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.

Citations for His Tag Antibody (HIS.H8) [HRP]

Customer Reviews for His Tag Antibody (HIS.H8) [HRP] (1)

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  • His Tag Antibody (HIS.H8)
    Name: Anonymous
    Application: Western Blot
    Sample Tested: His-tagged recombinant proteins
    Species: Human
    Verified Customer | Posted 08/22/2019
    Tested several different anti-his-HRP antibodies, this is the best, strong signal with no background and no non-specific bands in western blot.
    His Tag Antibody (HIS.H8) [HRP] NBP2-31055H

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Protocols

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