Recombinant
RabMAb

Recombinant Anti-6X His tag® antibody [JPAR-2] - BSA and Azide free (ab245203)

Overview

  • Product name

    Anti-6X His tag® antibody [JPAR-2] - BSA and Azide free
    See all 6X His tag® primary antibodies
  • Description

    Rabbit monoclonal [JPAR-2] to 6X His tag® - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, IHC, Flow Cyt, WBmore details
    Unsuitable for: ICC/IF
  • Species reactivity

    Reacts with: Species independent
  • Immunogen

    Synthetic peptide within 6X His tag® conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.

  • Positive control

    • WB: HEK-293T transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5; aa 1-1053; 125kDa) construct, whole cell lysate. IHC-P: HEK-293T transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5) construct. Flow cyt: HEK-293T transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5) construct.
  • General notes

    Ab245203 is the carrier-free version of ab245114. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab245203 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    JPAR-2
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab245203 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IP Use at an assay dependent concentration.
IHC Use at an assay dependent concentration.

Agarose-embedded sections.

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Flow Cyt Use at an assay dependent concentration.
WB Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for ICC/IF.
  • Target

    • Relevance

      The H-H-H-H-H-H motif is used as a tag on many recombinant proteins to facilitate purification. The antibody recognizes the His-tag fused to the amino- or carboxy- termini of targeted proteins in transfected or transformed cells.
    • Cellular localization

      Depends upon the localization of the parent protein tagged with hexahistidine.
    • Alternative names

      • 6 His epitope tag antibody
      • Hexa His tag antibody
      • HHHHHH epitope tag antibody
      • HHHHHH tag antibody
      • His tag antibody
      • Polyhistidine Tag antibody
      see all

    Images

    • His-tagged Staphylococcus aureus cas9 (J7RUA5; aa 1-1053; 125kDa) was immunoprecipitated from 0.35 mg of HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) (transfected with Staphylococcus aureus cas9 (J7RUA5; aa 1-1053; 125kDa)) whole cell lysate with ab245114 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab245114 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

      Lane 1: HEK-293T transfected with a His-tagged Staphylococcus aureus cas9 whole cell lysate 10 μg (Input).
      Lane 2: ab245114 IP in HEK-293T transfected with a His-tagged Staphylococcus aureus cas9 whole cell lysate. 
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab245114 in HEK-293T transfected with a His-tagged Staphylococcus aureus cas9 whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.
      Exposure time: 30 seconds.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab245114).

    • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5) construct labeling 6X His tag® with ab245114 at 1/500 (Right) compared with a Rabbit monoclonal IgG isotype control  (ab172730) (Left). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (ab150079), at 1/2000 dilution was used as the secondary antibody.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab245114).

    • Immunohistochemical analysis of agarose-embedded HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) labeling 6X His tag® with ab245114 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on (A) HEK-293T transfected with a 6× His tag construct is observed. No staining on (B) HEK-293T transfected with an empty plasmid is observed. Counter stained with hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab245114).

    References

    ab245203 has not yet been referenced specifically in any publications.

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    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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