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) [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) [NBP2-31055] - LEFT: untransfected control; RIGHT: anti-His (in red) on His-tagged fusion proteins in HEK293 cells. Both counterstained with DAPI (in blue).

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

Western Blot: His Tag Antibody (HIS.H8) [NBP2-31055] - Analysis using the DyLight 680 conjugate of NBP2-31055. Detection 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.

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

Western Blot: His Tag Antibody (HIS.H8) [NBP2-31055] - Standard ladder containing five different His-tagged proteins; untransfected control (1), HEK293 cells transfected with His-tagged protein vector (2).

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

Western Blot: His Tag Antibody (HIS.H8) [NBP2-31055] - Comparison between anti-His tag (HIS.H8 / EH158) mAb with 2 different vendor Abs, probed against a standard ladder containing five different His-tagged proteins. All Ab dilutions are 1:2000 (0.5 ug/mL).

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

Western Blot: His Tag Antibody (HIS.H8) [NBP2-31055] - Analysis of recombinant Treponema denticola major outer sheath His-tagged protein fragments (rMSP, rMSP-C, rMSP-V, rMSP-N) purified from E. coli was performed by loading 5 ug of per well. 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) [NBP2-31055] - 6-Histidine Epitope Tag Antibody (HIS.H8) [NBP2-31055] - Analysis using the DyLight 488 conjugate of NBP2-31055. Staining 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 488-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 Scietific ArrayScan or ToxInsight Instrument at 20X magnification.

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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.

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

Download Product Datasheets

COA

SDS

Download SDS

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

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

Key Product Details

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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]

Immunocytochemistry/ Immunofluorescence: 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)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)Azide and BSA Free [NBP2-31055]

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

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

ELISA

Flow Cytometry

Immunocytochemistry/ Immunofluorescence

Immunoprecipitation

Western Blot

Formulation, Preparation, and Storage

Purification

Formulation

Format

Preservative

Concentration

Shipping

Stability & Storage

Background: His Tag

Long Name

Alternate Names

Additional His Tag Products

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

Product Datasheets

COA

Certificate of Analysis

SDS

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