His Tag Antibody (HIS.H8) [DyLight 680]

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

Novus Biologicals, part of Bio-Techne
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Key Product Details

Species Reactivity

Epitope Tag

Applications

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

Label

DyLight 680 (Excitation = 692 nm, Emission = 712 nm)

Antibody Source

Monoclonal Mouse IgG2B Clone # HIS.H8

Concentration

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

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) [DyLight 680]

Western Blot: 6-Histidine Epitope Tag Antibody (HIS.H8) [DyLight 680] [NBP2-31055FR] - 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 a DyLight 680-conjugated 6x-His Epitope Tag monoclonal antibody at a dilution of 1:1000 for 1 hour at room temperature on a rocking platform and washed in TBS-0.1% Tween-20. Detection was performed using the LI-COR Odyssey.
Immunocytochemistry/Immunofluorescence: 6-Histidine Epitope Tag Antibody (HIS.H8) [DyLight 680] [NBP2-31055FR] - analysis of HeLa cells transfected with a construct containing a 6x-His Epitope Tag. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with a DyLight 680-conjugated 6x-His Epitope Tag monoclonal antibody at a dilution of 1:25 for at least 1 hour at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye. Images were taken on a Thermo Scientific ArrayScan or ToxInsight Instrument at 20X magnification.

Applications for His Tag Antibody (HIS.H8) [DyLight 680]

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.

Simple Western

Optimal dilutions of this antibody should be experimentally determined.

Western Blot

Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

50mM Sodium Borate

Preservative

0.02% Sodium Azide

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) [DyLight 680]

Product Datasheets

Certificate of Analysis

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Product Specific Notices for His Tag Antibody (HIS.H8) [DyLight 680]



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

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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