His Tag Antibody (AD1.1.10) - Non-Recombinant Monoclonal

Simple Western: His Tag Antibody (AD1.1.10) [NB100-64768] - Image shows a specific band for 6-His Epitope Tag in 0.1 mg/mL of rhEDG6 CHO S Tx lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Western Blot: His Tag Antibody (AD1.1.10) [NB100-64768] - Analysis using the Biotin conjugate of NB100-64768. Detection of HIS tagged protein probed with mouse anti histidine tag : Biotin and visualised with Streptavidin : HRP.

Flow Cytometry: His Tag Antibody (AD1.1.10) [NB100-64768] - Analysis using the FITC conjugate of NB100-64768. Staining of Human anti-human CD5 (HuCAL dHLX antibody with HIS tag) human peripheral blood lymphocytes, visualised with FITC conjugated Mouse anti-HIS tag antibody

Western Blot: His Tag Antibody (AD1.1.10) [NB100-64768] - Drosophila, 20 ug whole cell lysate

Western Blot: His Tag Antibody (AD1.1.10) [NB100-64768] - Analysis using the HRP conjugate of NB100-64768. Detection of His-Ubiquitin recombinant protein probed with Mouse anti Histidine Tag.

Western Blot: His Tag Antibody (AD1.1.10) [NB100-64768] - Analysis of 1) 6x-His tagged FLCN protein and 2) 6x-His tagged homer protein.

• Reactivity: ET
• Applications: WB, Simple Western, ELISA, Flow, Func, IHC, IP
• Clone: AD1.1.10
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: Non-Recombinant Monoclonal
• Concentration: 1.0 mg/ml

His Tag Antibody (AD1.1.10) - Non-Recombinant Monoclonal Summary

• Description: Novus Biologicals Mouse His Tag Antibody (AD1.1.10) - Non-Recombinant Monoclonal (NBP3-07011) is a monoclonal antibody validated for use in IHC, WB, ELISA, Flow, ICC/IF, Simple Western and IP. Anti-His Tag Antibody: Cited in 5 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Immunogen: This His Tag Antibody (AD1.1.10) - Non-Recombinant Monoclonal was developed against PAX6 transcription factor linked to histidine tag.
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • ELISA 1:100 - 1:500
  • Flow Cytometry 1:10 - 1:50
  • Immunohistochemistry 1:100 - 1:300
  • Immunohistochemistry-Frozen 1:100 - 1:300
  • Immunoprecipitation 1:10 - 1:500
  • In vitro assay
  • Simple Western 1:250
  • Western Blot 1:1000
• Application Notes: This antibody is routinely tested in WB using Histidine-tagged recombinant proteins. In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
  See Simple Western Antibody Database for Simple Western validation: Tested in rhEDG6 CHO S Tx lysate 0.1 mg/mL, Recombinant EGFP and rhSNAP25-GFP lysate 1.25 mg/mL; separated by Size; antibody dilution of 1:250, 1:50. Separated by Size-Wes, Sally Sue/Peggy Sue.

Reactivity Notes

6x His Tag specific

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for His Tag Antibody (AD1.1.10) - Non-Recombinant Monoclonal

• 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 (NB100-64768)(5)

Publications using NB100-64768

• Jang S, Song J, Kim N et al. Development of an antibody-like T-cell engager based on VH-VL heterodimer formation and its application in cancer therapy Biomaterials 2021-03-28 [PMID: 33774526]
  
  Details:
  Citation using the Non-Recombinant Monoclonal format of this antibody.
• Davids BJ, Liu CM, Hanson EM et al. Identification of Conserved Candidate Vaccine Antigens in the Surface Proteome of Giardia lamblia Infect Immun 2019-04-10 [PMID: 30962402] (WB, Human)
  
  Details:
  Citation using the Non-Recombinant Monoclonal format of this antibody.
• Puerta-Guardo H, Glasner DR, Espinosa DA et al. Flavivirus NS1 Triggers Tissue-Specific Vascular Endothelial Dysfunction Reflecting Disease Tropism Cell Rep 2019-02-07 [PMID: 30726741] (ICC/IF)
  
  Details:
  Citation using the Alexa Fluor 647 format of this antibody.
• Kim MJ, Vargas MR, Harlan BA et al. Nitration and Glycation Turn Mature NGF into a Toxic Factor for Motor Neurons: A Role for p75(NTR) and RAGE Signaling in ALS Antioxid Redox Signal 2017-05-25 [PMID: 28537420] (IP, WB, Mouse)
  
  Details:
  Citation using the Non-Recombinant Monoclonal format of this antibody.
• Tseng FJ, Liu YK, Chung YS et al. A fusion protein composed of receptor binding domain of vascular endothelial growth factor-A and constant region fragment of antibody: angiogenesis antagonistic activity Cytotechnology 2011-04-05 [PMID: 21461946] (ELISA)
  
  Details:
  Citation using the Non-Recombinant Monoclonal format of this antibody.

Reviews for His Tag Antibody (NB100-64768) (2)

Average Rating: 5

(Based on 2 reviews)

We have 2 reviews tested in 1 species: Drosophila.

reviewed by: Verified Customer WB Drosophila 07/10/2017

Summary

• Application: Western Blot
• Sample Tested: S2 lysate (with overexpressed recombinant protein)
• Species: Drosophila
• Lot: D-1

reviewed by: Eric Wang WB Drosophila 04/13/2017

Summary

• Application: Western Blot
• Sample Tested: 20 ug whole cell lysate
• Species: Drosophila
• Lot: c-4