His Tag Antibody

Recombinant Monoclonal Antibody
Catalog # Availability Size / Price Qty
MAB050R-100
MAB050R-SP
MAB050R-500
MAB050R-01M
Detection  of His Tag by Western Blot.
2 Images
Product Details
FAQs
Supplemental Products
Reviews (7)

His Tag Antibody Summary

Specificity
Detects proteins containing accessible consecutive histidine regions.  The antibody detects His tags located at the amino- or carboxyl- terminus.
Source
Recombinant Monoclonal Mouse IgG1 Clone # AD1.1.10R
Purification
Protein A or G purified from cell culture supernatant
Immunogen
His-tagged peptide
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.2 µg/mL
See below
Flow Cytometry
0.25 µg/106 cells
See below
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.
 

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.

Data Examples

Western Blot Detection  of His Tag by Western Blot. View Larger

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.

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

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

Reconstitution Calculator

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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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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
6 His epitope tag; 6X His; 6X-His; H; HHHHHH epitope tag; HHHHHH tag; His Tag; HIS; polyHistidine; Poly-histidine

Product Datasheets

FAQs

  1. 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.

View all Antibody FAQs

Isotype Controls

Mouse IgG1 Isotype Control

Ctrl

Reconstitution Buffers

Secondary Antibodies

Reviews for His Tag Antibody

Average Rating: 5 (Based on 7 Reviews)

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His Tag Antibody
By Anonymous on 08/26/2018
Application: Flow Sample Tested: B16-F1 mouse melanoma cell line Species: Mouse

His Tag Antibody
By Anonymous on 04/12/2018
Application: Flow Sample Tested: B16-F10 mouse melanoma cell line Species: Mouse

His Tag Antibody
By Anonymous on 03/30/2018
Application: Flow Sample Tested: B16-F1 mouse melanoma cell line Species: Mouse

His Tag Antibody
By Anonymous on 03/15/2018
Application: Flow Sample Tested: B16-F10 mouse melanoma cell line Species: Mouse

His Tag Antibody
By Anonymous on 03/11/2018
Application: Flow Sample Tested: 786-O human renal cell adenocarcinoma cell line Species: Mouse

His Tag Antibody
By Anonymous on 12/18/2017
Application: Flow Sample Tested: MCF 10A human breast epithelial cell line Species: Human

His Tag Antibody
By Anonymous on 11/27/2017
Application: Flow Sample Tested: B16-F1 mouse melanoma cell line Species: Mouse