His Tag Antibody (HIS.H8) - Azide and BSA Free

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-31055

Novus Biologicals, part of Bio-Techne

Key Product Details

Species Reactivity

Epitope Tag

Applications

Validated:

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

Cited:

Immunoprecipitation, Multiplex Immunoassay

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # HIS.H8

Format

Azide and BSA Free

Concentration

1 mg/ml

View all His Tag 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) - Azide and BSA Free

Western Blot: His Tag Antibody (HIS.H8)Azide and BSA Free [NBP2-31055]

Western Blot: His Tag Antibody (HIS.H8)Azide and BSA Free [NBP2-31055]

Western Blot: His Tag Antibody (HIS.H8) [NBP2-31055] - Analysis of various amounts of E. coli lysate containing a multi-epitope tagged protein per well.
Western Blot: His Tag Antibody (HIS.H8)Azide and BSA Free [NBP2-31055]

Western Blot: His Tag Antibody (HIS.H8)Azide and BSA Free [NBP2-31055]

Western Blot: His Tag Antibody (HIS.H8) [NBP2-31055] - Analysis of His-tagged protein in E. coli lysates. Western blot image submitted by a verified customer review.
Immunocytochemistry/ Immunofluorescence: His Tag Antibody (HIS.H8) - Azide and BSA Free [NBP2-31055]

Immunocytochemistry/ Immunofluorescence: His Tag Antibody (HIS.H8) - Azide and BSA Free [NBP2-31055]

Immunocytochemistry/Immunofluorescence: His Tag Antibody (HIS.H8) [NBP2-31055] - LEFT: untransfected control; RIGHT: anti-His (in red) on His-tagged fusion proteins in HEK293 cells. Both counterstained with DAPI (in blue).
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Applications for His Tag Antibody (HIS.H8) - Azide and BSA Free

Application
Recommended Usage

ELISA

1:100 - 1:2000

Flow Cytometry

1:10 - 1:1000

Immunocytochemistry/ Immunofluorescence

1:500 - 1:2000

Immunoprecipitation

1:10 - 1:500

Western Blot

1:1000 - 1:3000
Application Notes
Simple Western reported by an internal validation. Separated by Size-Jess/Wes, Sally Sue/Peggy Sue, antibody dilution of 1:50, 1:100; matrix was 12-230 kDa. See Simple Western Antibody Database for additional Simple Western validation: Tested in transfected cells; separated by size; antibody dilution of 1:100.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS (pH 7.2)

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

1 mg/ml

Shipping

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

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

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) - Azide and BSA Free

Product Datasheets

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

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Product Specific Notices for His Tag Antibody (HIS.H8) - Azide and BSA Free

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.

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