RecombinantRabMAb

Recombinant Anti-6X His tag® antibody [JPAR-2] (ab245114)

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Immunoprecipitation - Anti-6X His tag® antibody [JPAR-2] (ab245114)
  • Flow Cytometry - Anti-6X His tag® antibody [JPAR-2] (ab245114)
  • Immunohistochemistry - Anti-6X His tag® antibody [JPAR-2] (ab245114)
  • Western blot - Anti-6X His tag® antibody [JPAR-2] (ab245114)
  • Anti-6X His tag® antibody [JPAR-2] (ab245114)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [JPAR-2] to 6X His tag®
  • Suitable for: IHC, WB, IP, Flow Cyt
  • Reacts with: Species independent

Overview

  • Product name

    Anti-6X His tag® antibody [JPAR-2]
    See all 6X His tag® primary antibodies

  • Description

    Rabbit monoclonal [JPAR-2] to 6X His tag®

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC, WB, IP, Flow Cytmore 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

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • 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 ab245114 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
IHC 1/500.

Agarose-embedded sections.

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
WB 1/1000.
IP 1/30.
Flow Cyt 1/500.
  • 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

    • Immunoprecipitation - Anti-6X His tag® antibody [JPAR-2] (ab245114)
      Immunoprecipitation - Anti-6X His tag® antibody [JPAR-2] (ab245114)

      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.

    • Flow Cytometry - Anti-6X His tag® antibody [JPAR-2] (ab245114)
      Flow Cytometry - Anti-6X His tag® antibody [JPAR-2] (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.

    • Immunohistochemistry - Anti-6X His tag® antibody [JPAR-2] (ab245114)
      Immunohistochemistry - Anti-6X His tag® antibody [JPAR-2] (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. Counterstained 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).

    • Western blot - Anti-6X His tag® antibody [JPAR-2] (ab245114)
      Western blot - Anti-6X His tag® antibody [JPAR-2] (ab245114)
      All lanes : Anti-6X His tag® antibody [JPAR-2] (ab245114) at 1/1000 dilution

      Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with an empty plasmid, whole cell lysate
      Lane 2 : HEK-293T transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5; aa 1-1053; 125kDa) construct, whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Observed band size: 125 kDa why is the actual band size different from the predicted?


      Exposure time: 15 seconds


      Blocking and dilution buffer: 5% NFDM/TBST.

    • Anti-6X His tag® antibody [JPAR-2] (ab245114)
      Anti-6X His tag® antibody [JPAR-2] (ab245114)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (0)

    Publishing research using ab245114? Please let us know so that we can cite the reference in this datasheet.

    ab245114 has not yet been referenced specifically in any publications.

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