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

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) [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) [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) [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) [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) [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) [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) [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.

• Reactivity: ET
• Applications: WB, Simple Western, ELISA, Flow, ICC/IF, IP, CyTOF-ready
• Clone: HIS.H8
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: Azide and BSA Free
• Concentration: 1 mg/ml

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

• Description: Novus Biologicals Mouse His Tag Antibody (HIS.H8) - Azide and BSA Free (NBP2-31055) is a monoclonal antibody validated for use in WB, ELISA, Flow, ICC/IF, Simple Western and IP. Anti-His Tag Antibody: Cited in 3 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Immunogen: This His Tag Antibody (HIS.H8) was developed against a 6x His synthetic peptide.
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • ELISA 1:100 - 1:2000
  • Flow Cytometry 1:10 - 1:1000
  • Immunocytochemistry/ Immunofluorescence 1:500 - 1:2000
  • Immunoprecipitation 1:10 - 1:500
  • Simple Western
  • Western Blot 1:1000 - 1:3000
• Application Notes: See Simple Western Antibody Database for Simple Western validation: Tested in Transfected cells, separated by Size, antibody dilution of 1:100

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS (pH 7.2)
• Preservative: No Preservative
• Concentration: 1 mg/ml
• Purity: Protein A purified

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

• 6 His epitope tag
• 6-His Tag
• 6X His Tag
• 6X His
• 6x-His Tag
• 6X-His
• H
• Hexa His tag
• HHHHHH epitope tag
• HHHHHH tag
• His Tag
• HIS
• His6-Tag
• polyhistidine tag
• polyHistidine
• Poly-histidine

Background

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

Limitations

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.

Publications for His Tag Antibody (NBP2-31055)(3)

Publications using NBP2-31055

• Thompson VF, Wieland DR, Mendoza-Leon V et al. Binding of the nuclear ribonucleoprotein family member FUS to RNA prevents R-loop RNA:DNA hybrid structures The Journal of biological chemistry 2023-09-08 [PMID: 37690693] (IP)
• Ihara S, Miyamoto Y, Le CHY et al. Conserved metabolic enzymes as vaccine antigens for giardiasis PLoS neglected tropical diseases [PMID: 35468132] (MI)
• Ajam T, De I, Petkau N Et al. Alternative catalytic residues in the active site of Esco acetyltransferases Sci Rep 2020-06-20 [PMID: 32555289]
  
  Details:
  Citation using the HRP version of this antibody.