Key Product Details

Species Reactivity

Validated:

Multi-Species

Cited:

Human, E. coli, Yeast

Applications

Validated:

Western Blot, Flow Cytometry, Immunoaffinity Purification

Cited:

Western Blot, Neutralization

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Mouse IgG1 Clone # AD1.1.10R
View all His Tag Primary Antibodies »

Product Specifications

Immunogen

His-tagged peptide

Specificity

Detects proteins containing accessible consecutive histidine regions.  The antibody detects His tags located at the amino- or carboxyl- terminus.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for His Tag Antibody

Detection of His Tag antibody by Western Blot.

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.

Detection of His Tag antibody in HEK293 Human Cell Line Transfected with His-tagged Proteins antibody by Flow Cytometry.

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.

Detection of His Tag by Western Blot

Detection of His Tag by Western Blot

Lactate directly binds & inhibits the activity of PHD2. d His-tagged HIF-1 alpha was incubated with PHD2 in presence of 20 mM lactate or NaCl. Time-dependent HIF-1 alpha hydroxylation was examined by WBting. Image collected & cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36064857), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of His Tag by Western Blot

Detection of His Tag by Western Blot

Lactate directly binds & inhibits the activity of PHD2. d His-tagged HIF-1 alpha was incubated with PHD2 in presence of 20 mM lactate or NaCl. Time-dependent HIF-1 alpha hydroxylation was examined by WBting. Image collected & cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36064857), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for His Tag Antibody

Application
Recommended Usage

Flow Cytometry

0.25 µg/106 cells
Sample: HEK293 human embryonic kidney cell line transfected with His-tagged proteins

Immunoaffinity Purification

Immobilized anti-His antibody can be used to affinity purify proteins containing exposed His tags. The column capacity must be determined for each individual His-tagged protein.

Western Blot

0.2 µg/mL
Sample: HEK293 human embryonic kidney cell line and CHO Chinese hamster ovary cell line transfected with His-tagged proteins

Reviewed Applications

Read 8 reviews rated 5 using MAB050R in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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.

Long Name

Histidine Tag

Alternate Names

polyHistidine

Additional His Tag Products

Product Documents for His Tag Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for His Tag Antibody

For research use only

Citations for His Tag Antibody

Customer Reviews for His Tag Antibody (8)

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8 Customer Ratings
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Customer Images

Showing  1 - 5 of 8 reviews Showing All
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  • His Tag Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Human recombinant test antibody
    Species: Mouse
    Verified Customer | Posted 12/21/2018
  • His Tag Antibody
    Name: Anonymous
    Application: Flow Cytometry
    Sample Tested: B16-F1 mouse melanoma cell line
    Species: Mouse
    Verified Customer | Posted 08/26/2018
  • His Tag Antibody
    Name: Anonymous
    Application: Flow Cytometry
    Sample Tested: B16-F10 mouse melanoma cell line
    Species: Mouse
    Verified Customer | Posted 04/12/2018
  • His Tag Antibody
    Name: Anonymous
    Application: Flow Cytometry
    Sample Tested: B16-F1 mouse melanoma cell line
    Species: Mouse
    Verified Customer | Posted 03/30/2018
  • His Tag Antibody
    Name: Robert Reddy
    Application: Flow Cytometry
    Sample Tested: B16-F10 mouse melanoma cell line
    Species: Mouse
    Verified Customer | Posted 03/15/2018
  • His Tag Antibody
    Name: Anonymous
    Application: Flow Cytometry
    Sample Tested: 786-O human renal cell adenocarcinoma cell line
    Species: Mouse
    Verified Customer | Posted 03/11/2018
  • His Tag Antibody
    Name: Amit Kumar
    Application: Flow Cytometry
    Sample Tested: MCF 10A human breast epithelial cell line
    Species: Human
    Verified Customer | Posted 12/18/2017
  • His Tag Antibody
    Name: Anonymous
    Application: Flow Cytometry
    Sample Tested: B16-F1 mouse melanoma cell line
    Species: Mouse
    Verified Customer | Posted 11/27/2017
    His Tag Antibody MAB050R

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FAQs for His Tag Antibody

Showing  1 - 3 of 3 FAQs Showing All

    • A: 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.

    • Q: 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?
      A: 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.

    • Q: What is the difference between His Tag Antibodies, Catalog #s MAB050 and MAB050R?
      A: 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.
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