His Tag Antibody Summary
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
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Detection of His Tag by Western Blot. Western blot shows lysates of HEK293 human embryonic kidney cell line either non-transfected or transfected with His-tagged human CIQ4 and human MYCOC and CHO Chinese hamster ovary cell line transfected with His-tagged human BAI-1. PVDF membrane was probed with 0.2 µg/mL of Mouse Anti-His Tag Recombinant Monoclonal Antibody (Catalog # MAB050R) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). Specific bands were detected for His Tag at approximately 32, 65, and 120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
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Detection of His Tag in HEK293 Human Cell Line Transfected with His-tagged Proteins by Flow Cytometry. HEK293 human embryonic kidney cell line transfected with His-tagged proteins were stained with Mouse Anti-His Tag Recombinant Monoclonal Antibody (Catalog # MAB050R, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.
Reconstitution Calculator
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: His Tag
Consecutive histidine residues (usually 6 to 10 in length) are often inserted into the amino acid sequences of recombinant proteins. The resulting His-tagged proteins can be detected or purified by using anti-polyHis antibodies.
Product Datasheets
Citations for His Tag Antibody
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 6
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folA thyA knockout E. coli as a suitable surrogate model for evaluation of antifolate sensitivity against PfDHFR-TS
Authors: Suwanakitti, N;Talawanich, Y;Vanichtanankul, J;Taweechai, S;Yuthavong, Y;Kamchonwongpaisan, S;Kongkasuriyachai, D;
Acta tropica
Species: E. coli
Sample Types: Cell Lysates
Applications: Western Blot -
Synonymous mutations in the phosphoglycerate kinase 1 gene induce an altered response to protein misfolding in Schizosaccharomyces pombe
Authors: S Moreira-Ra, L Arias, R Flores, A Katz, G Levicán, O Orellana
Frontiers in Microbiology, 2023-01-11;13(0):1074741.
Species: Yeast
Sample Types: Protein
Applications: Western Blot -
Adipocyte-derived lactate is a signalling metabolite that potentiates adipose macrophage inflammation via targeting PHD2
Authors: T Feng, X Zhao, P Gu, W Yang, C Wang, Q Guo, Q Long, Q Liu, Y Cheng, J Li, CKY Cheung, D Wu, X Kong, Y Xu, D Ye, S Hua, K Loomes, A Xu, X Hui
Nature Communications, 2022-09-05;13(1):5208.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
Paradoxical roles of ATF6α and ATF6β in modulating disease severity caused by mutations in collagen X.
Authors: Forouhan M, Mori K, Boot-Handford R
Matrix Biol, 2018-03-06;0(0):.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
Lipid bilayer simulations of CXCR4 with inverse agonists and weak partial agonists.
Authors: Trent J, Wang Z, Murray J, Shao W, Tamamura H, Fujii N, Peiper S
J Biol Chem, 2003-09-04;278(47):47136-44.
Applications: Western Blot -
Modification of Transfer RNA Levels Affects Cyclin Aggregation and the Correct Duplication of Yeast Cells
Authors: Loreto Arias, Fabián Martínez, Daniela González, Rodrigo Flores-Ríos, Assaf Katz, Mario Tello et al.
Frontiers in Microbiology
FAQs
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This protein datasheet indicates I need to use a cross-linking antibody, Catalog # MAB050, for biological activity. What is this antibody and is it really necessary?
The antibody is directed against a 6x histidine repeat and is recommended for use as a cross-linker of proteins with 6x his-tag. Crosslinking is often used for proteins that require receptor trimerization and can result greater biological activity. R&D Systems Quality Control tests the performance of these proteins in the presence of the cross-linking antibody. Therefore, it is necessary to use this antibody when trying to achieve the same level of specific activity described in the datasheet.
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Can the His Tag Antibody (MAB050) recognize all His-tagged proteins?
MAB050 has been validated specifically for the detection of His-tags localized to the amino- or carboxyl-terminus of a peptide. It is also expected to detect any series of 6 or more histidine residues as long as this epitope is conformationally accessible. For applications utilizing linearized proteins, it is likely that MAB050 will have access to any consecutive histidine regions. If used with protein in its native conformation, there is the potential that the protein will be folded such that the antibody is not able to bind its target.
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What is the difference between His Tag Antibodies, Catalog #s MAB050 and MAB050R?
The primary difference is how the antibodies are produced. Catalog # MAB050R is made using recombinant DNA technology while Catalog # MAB050 is produced from a hybridoma.
Reviews for His Tag Antibody
Average Rating: 5 (Based on 8 Reviews)
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