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His Tag Antibody

Catalog # MAB050 | R&D Systems, Inc. a Bio-Techne Brand
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MAB050-100
MAB050-500
MAB050-SP

Key Product Details

Species Reactivity

Multi-Species

Applications

CyTOF-ready, Flow Cytometry, Immunoaffinity Purification, Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # AD1.1.10

Product Summary for His Tag Antibody

Immunogen

His-tagged peptide

Specificity

Detects proteins containing accessible consecutive histidine regions. The antibody detects His tags localized 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 Monoclonal Antibody (Catalog # MAB050) 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 was stained with Mouse Anti-His Tag Monoclonal Antibody (Catalog # MAB050, 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).

SARS-Cov-2 Spike 1 His tag protein binding to ACE-2-transfected Human Cell Line is detected by His tag Antibody.

Recombinant SARS-Cov-2 Spike 1 His-tagged protein (10522-CV) binds to HEK293 human embryonic kidney cell line transfected with recombinant human ACE-2 and was detected with Mouse Anti-His Allophycocyanin-conjugated Monoclonal Antibody (IC050A, filled histogram). No staining was observed in the absence of protein (open histogram). Staining was performed using our Staining Membrane-Associated Proteins protocol.

Applications for His Tag Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/106 cells
Sample: HEK293 human embryonic kidney cell line transfected with His-tagged proteins and SARS-Cov-2 Spike 1 His tag protein binding to ACE-2-transfected Human Cell Line

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.

Simple Western

10 µg/mL
Sample: CHO Chinese hamster ovary cell line transfected with His-tagged Human S1P4/EDG-6

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
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 11 reviews rated 4.9 using MAB050 in the following applications:

Formulation, Preparation and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Endotoxin Level

<0.20 EU per 1 μg of the antibody by the LAL method.

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

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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

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C

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.

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

Product Documents for His Tag Antibody

Certificate of Analysis

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

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for His Tag Antibody

For research use only

Citations for His Tag Antibody (57)

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Customer Reviews for His Tag Antibody (11)

4.9 out of 5
11 customer ratings
5 stars
91%
4 stars
9%
3 stars
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1 star
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  • His Tag Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: 293T human embryonic kidney cell line
    Species: Human
    Verified Customer | Posted 04/06/2022
    His Tag Antibody MAB050
  • His Tag Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: HEK293T human embryonic kidney cell line
    Species: Human
    Verified Customer | Posted 10/29/2021
    His Tag Antibody MAB050
  • His Tag Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Recombinant protein
    Species: Mouse
    Verified Customer | Posted 08/06/2021
    His Tag Antibody MAB050
  • His Tag Antibody
    Name: Jenna Nguyen
    Application: Flow Cytometry
    Sample Tested: 7TD1 mouse hybridoma cell line
    Species: Mouse
    Verified Customer | Posted 05/07/2019
  • His Tag Antibody
    Name: Jenna Nguyen
    Application: Cell depletion
    Sample Tested: 4T1 mouse breast cancer cell line
    Species: Mouse
    Verified Customer | Posted 01/11/2019
  • His Tag Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: 293T human embryonic kidney cell line
    Species: Human
    Verified Customer | Posted 07/23/2018
    293T cells were transfected with pCDNA 3.1-Control or His-RIP1 for 48h. Total cell lysates were subjected to western blot. PVDF membrane were probed with 1 um/ml Human His Tag Antibody (MAB050). A specific band was detected for His-RIP1 at approximately 75 kDa. This experiment was conducted under reducing conditions
    His Tag Antibody MAB050
  • His Tag Antibody
    Name: Anonymous
    Application: Simple Western
    Sample Tested: Purified protein
    Species: Human
    Verified Customer | Posted 06/20/2018
    His Tag Antibody MAB050
  • His Tag Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: 293T human embryonic kidney cell line
    Species: Mouse
    Verified Customer | Posted 07/15/2017
    His Tag Antibody MAB050
  • His Tag Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: HEK293T human embryonic kidney cell line
    Species: Human
    Verified Customer | Posted 06/28/2016
    cell lysates transformed
  • His Tag Antibody
    Name: Rachel Jacobson
    Application: Western Blot
    Sample Tested: Recombinant protein
    Species: Human
    Verified Customer | Posted 06/28/2016
    His Tag Antibody MAB050
  • His Tag Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: Human recombinant test antibody
    Species: Human
    Verified Customer | Posted 10/26/2015
    Specificity: Specific
    Sensitivity: Sensitive
    Buffer: TBS + Tween + BSA
    Dilution: used at 2ug/mL

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FAQs for His Tag Antibody

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  • Q: Can the His Tag Antibody (MAB050) recognize all His-tagged proteins?

    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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