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His Tag Antibody
R&D Systems, Inc. a Bio-Techne Brand
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Key Product Details
| Immunogen: |
His-tagged peptide |
| Applications: | Affinity Purification, Flow Cytometry, Western Blot |
| Label: | Unconjugated |
| Source: | Recombinant Monoclonal Mouse IgG1 Clone # AD1.1.10R |
| Specificity: |
Detects proteins containing accessible consecutive histidine regions. The antibody detects His tags located at the amino- or carboxyl- terminus. |
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Western BlotDetection 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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Flow CytometryDetection 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. |
Citations for His Tag Antibody (4)
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Citations are publications that use Bio-Techne products. Selected citations for His Tag Antibody include:
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Species: Human
Sample Types: Cell Lysates
Applications: Western BlotT Feng et al. (2022), Adipocyte-derived lactate is a signalling metabolite that potentiates adipose macrophage inflammation via targeting PHD2 Nature Communications, 13(1):5208.
PMID: 36064857 -
Species: Yeast
Sample Types: Protein
Applications: Western BlotS Moreira-Ra et al. (2022), Synonymous mutations in the phosphoglycerate kinase 1 gene induce an altered response to protein misfolding in Schizosaccharomyces pombe Frontiers in Microbiology, 13(0):1074741.
PMID: 36713198 -
Species: Human
Sample Types: Cell Lysates
Applications: Western BlotForouhan M et al. (0), Paradoxical roles of ATF6α and ATF6β in modulating disease severity caused by mutations in collagen X. Matrix Bio0(0).
PMID: 29522813 -
Sample Types: Protein
Applications: Western BlotTrent J et al. (0), Lipid bilayer simulations of CXCR4 with inverse agonists and weak partial agonists. J Biol Che278(47):47136-44.
PMID: 12958314
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Customer Reviews for His Tag Antibody (8)
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8 customer ratings
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Application: WBSample Tested: Human recombinant test antibodySpecies: MouseVerified Customer | Posted 12/21/2018
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Application: FlowSample Tested: B16-F1 mouse melanoma cell lineSpecies: MouseVerified Customer | Posted 08/26/2018
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Application: FlowSample Tested: B16-F10 mouse melanoma cell lineSpecies: MouseVerified Customer | Posted 04/12/2018
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Application: FlowSample Tested: B16-F1 mouse melanoma cell lineSpecies: MouseVerified Customer | Posted 03/30/2018
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Application: FlowSample Tested: B16-F10 mouse melanoma cell lineSpecies: MouseVerified Customer | Posted 03/15/2018
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Application: FlowSample Tested: 786-O human renal cell adenocarcinoma cell lineSpecies: MouseVerified Customer | Posted 03/11/2018
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Application: FlowSample Tested: MCF 10A human breast epithelial cell lineSpecies: HumanVerified Customer | Posted 12/18/2017
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Application: FlowSample Tested: B16-F1 mouse melanoma cell lineSpecies: MouseVerified Customer | Posted 11/27/2017
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FAQs for His Tag Antibody
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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.
