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RecombinantRabMAb

Recombinant Anti-WASP/Wiskott-Aldrich syndrome protein antibody [EP2541Y] (ab75830)

  • Datasheet
Submit a review Q&A (1)

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Immunocytochemistry - Anti-WASP/Wiskott-Aldrich syndrome protein antibody [EP2541Y] (ab75830)
  • Flow Cytometry - Anti-WASP/Wiskott-Aldrich syndrome protein antibody [EP2541Y] (ab75830)
  • Western blot - Anti-WASP/Wiskott-Aldrich syndrome protein antibody [EP2541Y] (ab75830)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP2541Y] to WASP/Wiskott-Aldrich syndrome protein
  • Suitable for: WB, IHC-P, Flow Cyt, ICC
  • Reacts with: Human

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Overview

  • Product name

    Anti-WASP/Wiskott-Aldrich syndrome protein antibody [EP2541Y]
    See all WASP/Wiskott-Aldrich syndrome protein primary antibodies
  • Description

    Rabbit monoclonal [EP2541Y] to WASP/Wiskott-Aldrich syndrome protein
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, ICCmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human WASP/Wiskott-Aldrich syndrome protein aa 450-550 (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: TF-1 and U-937 whole cell lysates; ICC: TF-1 cells; Flow Cyt: U-937 cells.
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     This product was previously labelled as WASP

     

    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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    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. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP2541Y
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Adapters
    • Cytoplasmic

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • U937 cytoplasmic extract lysate (ab14904)
  • Recombinant Protein

    • Recombinant Human WASP/Wiskott-Aldrich syndrome protein (ab152802)

Applications

Our Abpromise guarantee covers the use of ab75830 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
WB 1/10000 - 1/50000. Detects a band of approximately 60 kDa (predicted molecular weight: 53 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cyt 1/30.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

ICC 1/50.
  • Application notes
    Is unsuitable for IP.
  • Target

    • Function

      Effector protein for Rho-type GTPases, providing a link with the Arp2/3 complex that regulates the structure and dynamics of the actin cytoskeleton. Important for efficient actin polymerization. Possible regulator of lymphocyte and platelet function.
    • Tissue specificity

      Expressed predominantly in the thymus. Also found, to a much lesser extent, in the spleen.
    • Involvement in disease

      Defects in WAS are the cause of Wiskott-Aldrich syndrome (WAS) [MIM:301000]; also known as eczema-thrombocytopenia-immunodeficiency syndrome. WAS is an X-linked recessive immunodeficiency characterized by eczema, thrombocytopenia, recurrent infections, and bloody diarrhea. Death usually occurs before age 10.
      Defects in WAS are the cause of thrombocytopenia type 1 (THC1) [MIM:313900]. Thrombocytopenia is defined by a decrease in the number of platelets in circulating blood, resulting in the potential for increased bleeding and decreased ability for clotting.
      Defects in WAS are a cause of neutropenia severe congenital X-linked (XLN) [MIM:300299]. XLN is an immunodeficiency syndrome characterized by recurrent major bacterial infections, severe congenital neutropenia, and monocytopenia.
    • Sequence similarities

      Contains 1 CRIB domain.
      Contains 1 WH1 domain.
      Contains 1 WH2 domain.
    • Domain

      The WH1 (Wasp homology 1) domain may bind a Pro-rich ligand.
      The CRIB (Cdc42/Rac-interactive-binding) region binds to the C-terminal WH2 domain in the autoinhibited state of the protein. Binding of Rho-type GTPases to the CRIB induces a conformation change and leads to activation.
    • Cellular localization

      Cytoplasm > cytoskeleton.
    • Target information above from: UniProt accession P42768 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 7454 Human
      • Omim: 300392 Human
      • SwissProt: P42768 Human
      • Unigene: 2157 Human
      • Alternative names

        • Eczema thrombocytopenia antibody
        • IMD2 antibody
        • SCNX antibody
        • THC antibody
        • THC1 antibody
        • Thrombocytopenia 1 (X linked) antibody
        • U42471 antibody
        • Was antibody
        • WASp antibody
        • WASP_HUMAN antibody
        • Wiskott Aldrich syndrome (eczema thrombocytopenia) antibody
        • Wiskott Aldrich syndrome antibody
        • Wiskott Aldrich syndrome protein antibody
        • Wiskott-Aldrich syndrome protein antibody
        see all

      Images

      • Immunocytochemistry - Anti-WASP/Wiskott-Aldrich syndrome protein antibody [EP2541Y] (ab75830)
        Immunocytochemistry - Anti-WASP/Wiskott-Aldrich syndrome protein antibody [EP2541Y] (ab75830)

        Immunocytochemistry analysis of TF-1 (Human bone marrow erythroblast) cells labeling WASP/Wiskott-Aldrich syndrome protein with Purified ab75830 at 1:50 dilution (5.88 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor® 488,ab150077) was used as the secondary antibody at 1:1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

      • Flow Cytometry - Anti-WASP/Wiskott-Aldrich syndrome protein antibody [EP2541Y] (ab75830)
        Flow Cytometry - Anti-WASP/Wiskott-Aldrich syndrome protein antibody [EP2541Y] (ab75830)
        Flow Cytometry analysis of U-937 (Human histiocytic lymphoma monocyte) cells labeling WASP/Wiskott-Aldrich syndrome protein with Purified ab75830 at 1:30 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488,ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
      • Western blot - Anti-WASP/Wiskott-Aldrich syndrome protein antibody [EP2541Y] (ab75830)
        Western blot - Anti-WASP/Wiskott-Aldrich syndrome protein antibody [EP2541Y] (ab75830)
        All lanes : Anti-WASP/Wiskott-Aldrich syndrome protein antibody [EP2541Y] (ab75830) at 1/10000 dilution (Purified)

        Lane 1 : TF-1 (Human Erythroleukemia erythroblast) whole cell lysate
        Lane 2 : U-937 (Human histiocytic lymphoma monocyte) whole cell lysate

        Lysates/proteins at 15 µg per lane.

        Secondary
        All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

        Predicted band size: 53 kDa
        Observed band size: 60 kDa
        why is the actual band size different from the predicted?



        Blocking buffer: 5% NFDM/TBST

      Protocols

      • Flow cytometry protocols
      • Immunohistochemistry protocols
      • Immunocytochemistry & immunofluorescence protocols
      • Western blot protocols

      Click here to view the general protocols

      Datasheets and documents

      • Datasheet
    • References (0)

      Publishing research using ab75830? Please let us know so that we can cite the reference in this datasheet.

      ab75830 has not yet been referenced specifically in any publications.

      Customer reviews and Q&As

      Show All Reviews Q&A
      Submit a review Submit a question

      Question

      Dear Sir or Madam!

      I would like to kindly ask some questions about your product:
      Code: ab75830
      Product name: Anti-WASP antibody [EP2541Y]
      Lot:892315

      I have read the datasheet of this product and you wrote this antibody is unsuitable for Flow cytometry. I would like to kindly ask you why you say that. Did you make some tests with this antibody and relying upon these findings you wrote it is unsuitable for Flow cytometry or you did not try this product in Flow cytometry analysis?
      I tried this WASP antibody by Flow cytometer and I got prosperous results.

      Please, send me more details with this problem, because I would like to use this antibody by flow cytometer but I read there are no references.

      (I also tried this product in Western blot analysis and I got similar results which I found by Flow cytometry. On the basis of this finding I thought it is suitable for Flow cytometry.)

      You wrote the concentration is not determined. But the concentration values are important data in Flow cytometry analysis. So I would like to kindly ask you about the concentration value in ug/ml.

      I would like to say thank you in advance.
      Yours faithfully,

      Read More

      Abcam community

      Verified customer

      Asked on Nov 06 2012

      Answer

      Thank you for your enquiry.

      I have contacted our laboratory source for this antibody. They have confirmed that the antibody was only tested once with one protocol in flow cytometry. In order to be honest with our customers, we have therefore stated does not work on the datasheet and we cannot guarantee it will work in flow cytometry until it is successfully tested. I am sorry we have no other flow cytometry data on this occasion. In this case, with a more thoroughly optimized protocol, it may be that the antibody will work.

      I am pleased to hear it has worked in your flow cytometry experiments. We like to encourage all of our customers to submit an Abreview via the online product datasheet, and this would be particularly beneficial in this case. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

      To find out more about our Abreview system, please see the following link: https://www.abcam.com/abreviews

      The originator of the antibody is also going to retest the antibody in this application. This may take some time, but I will be pleased to forward any additional data that we are sent.

      Should any further data we receive from the originator or in an Abreview from a customer indicate that this antibody will work in flow cytometry, we will be able to amend the datasheet to say successfully tested in flow cytometry.

      I hope this information is helpful. Please do not hesitate to contact me for any further advice or information in this regard.

      Read More

      Abcam Scientific Support

      Answered on Nov 06 2012

      Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
      For licensing inquiries, please contact partnerships@abcam.com

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