Key Product Details

Species Reactivity

Validated:

Epitope Tag

Cited:

Rat

Applications

Validated:

ELISA, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Western Blot

Cited:

IF/IHC, Immunocytochemistry/ Immunofluorescence, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Format

BSA Free

Concentration

1.0 mg/ml

View all His Tag Antibodies »

Product Specifications

Immunogen

This His Tag Antibody was developed against Keyhole limpet haemocyanin (KLH) conjugated H-H-H-H-H-H.

Specificity

This antibody recognizes the motif H-H-H-H-H-H and is therefore of value in detecting proteins containing histidine tags. This antibody may not detect all N-terminal (NT) histidine tagged proteins.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for His Tag Antibody - BSA Free

Western Blot: His Tag AntibodyBSA Free [NBP1-25939]

Western Blot: His Tag AntibodyBSA Free [NBP1-25939]

Western Blot: His Tag Antibody [NBP1-25939] - 200, 100, or 50 ng of E. coli whole cell lysate expressing a multi-tag fusion protein. Antibody used for WB at 0.2 ug/ml (1:5,000). Detection: Chemiluminescence withan exposure time of 3 minutes.

Applications for His Tag Antibody - BSA Free

Application
Recommended Usage

ELISA

1:1000 - 1:30000

Immunocytochemistry/ Immunofluorescence

1:100 - 1:400

Immunoprecipitation

1 - 4 ug/mg lysate

Western Blot

1:1000 - 1:5000
Application Notes
For coating plates in ELISA use antibody at 1:100 - 1:500.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.09% Sodium Azide

Concentration

1.0 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. 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 - 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 - 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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