His Tag Antibody (RM146)

Western Blot: His Tag Antibody (RM146) [NBP2-61482] - Western blot of 293T cells (1) untransfected or (2) transfected with His-Tag fusion protein X and (3) E. coli lysate without His-Tag Protein Y (4) E. coli lysate with His-Tag Protein Y, using NBP2-61482.

Immunohistochemistry: His Tag Antibody (RM146) [NBP2-61482] - Immunohistochemistry staining of naive HepG2 cells (Negative control) using anti-His-Tag antibody, NBP2-61482.

Immunohistochemistry-Paraffin: His Tag Antibody (RM146) [NBP2-61482] - Immunohistochemistry staining of 293T cells expressing His-Tag nuclear protein X, using NBP2-61482.

Simple Western: His Tag Antibody (RM146) [NBP2-61482] - Human purified protein. Simple Western image submitted by a verified customer review.

Immunoprecipitation: His Tag Antibody (RM146) [NBP2-61482] - Immunoprecipitation of 293T cells expressing a His-Tag fusion protein by NBP2-61482, was blotted with NBP2-61482. (1) Whole lysate control; (2) IP by rabbit IgG control; (3) IP by NBP2-61482.

• Reactivity: ET
• Applications: WB, Simple Western, Flow, ICC/IF, IHC, IP
• Clone: RM146
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

His Tag Antibody (RM146) Summary

• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: This His Tag Antibody (RM146) was developed against a mixture of a peptide with 6xHis-Tag at the N-terminus and another peptide with 6xHis-Tag at the C-terminus.
• Specificity: This antibody reacts to recombinant proteins containing the 6xHis-Tag or 10xHis-Tag fused to either the amino or carboxy terminus. No cross reactivity with other endogenous protein in mammalian or bacteria cells.
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 0.5 ug/mL - 2 ug/mL
  • Immunocytochemistry/ Immunofluorescence 0.5 ug/mL - 2 ug/mL
  • Immunohistochemistry 0.1 ug/mL - 1 ug/mL
  • Immunohistochemistry-Paraffin
  • Immunoprecipitation 0.5 ug/ml - 2 ug/ml
  • Simple Western reported from a verified customer review.
  • Western Blot 0.1 ug/ml - 1 ug/ml
• Application Notes: His Tag Antibody (RM146) validated for Simple Western from a verified customer review.
  See Simple Western Antibody Database for Simple Western validation: Tested in Human purified protein, separated by Size

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: 50% Glycerol/PBS, 1% BSA
• Preservative: 0.09% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for His Tag Antibody (RM146)

• 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 (NBP2-61482)(4)

Publications using NBP2-61482

• Hyo Young Jung, Hyun Jung Kwon, Kyu Ri Hahn, Woosuk Kim, Dae Young Yoo, Yeo Sung Yoon, Dae Won Kim, In Koo Hwang Tat-heat shock protein 10 ameliorates age-related phenotypes by facilitating neuronal plasticity and reducing age-related genes in the hippocampus Aging (Albany NY) 2023-11-30 [PMID: 38011257]
• Paul S, Fatihi S, Sharma S et al. Cyclin-Dependent Kinase 5 Regulates cPLA2 Activity and Neuroinflammation in Parkinson's Disease eNeuro 2022-11-23 [PMID: 36351818] (WB)
• Clement CC, Osan J, Buque A et al. PDIA3 epitope-driven immune autoreactivity contrWButes to hepatic damage in type 2 diabetes Science immunology 2022-08-12 [PMID: 35984892] (WB)
  
  Details:
  1:1000 dilution in PBS
• JF Arboleda-V, F Lopera, M O'Hare, S Delgado-Ti, C Marino, N Chmielewsk, KL Saez-Torre, D Amarnani, AP Schultz, RA Sperling, D Leyton-Cif, K Chen, A Baena, D Aguillon, S Rios-Romen, M Giraldo, E Guzmán-Vél, DJ Norton, E Pardilla-D, A Artola, JS Sanchez, J Acosta-Uri, M Lalli, KS Kosik, MJ Huentelman, H Zetterberg, K Blennow, RA Reiman, J Luo, Y Chen, P Thiyyagura, Y Su, GR Jun, M Naymik, X Gai, M Bootwalla, J Ji, L Shen, JB Miller, LA Kim, PN Tariot, KA Johnson, EM Reiman, YT Quiroz Resistance to autosomal dominant Alzheimer's disease in an APOE3 Christchurch homozygote: a case report Nat. Med., 2019-11-04;0(0):. 2019-11-04 [PMID: 31686034]

Review for His Tag Antibody (NBP2-61482) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Verified Customer Simple Western Human 12/04/2018

Summary

• Application: Simple Western
• Sample Tested: Purified protein
• Species: Human
• Lot: R-09-01742