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Signal Transduction Protein Phosphorylation Ser / Thr Kinases MAPK Pathway
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Validated using a knockout cell lineRabMAb

Anti-Crk p38 antibody [EP242Y] (ab45136)

  • Datasheet
Reviews (1) Submit a question References (1)

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Western blot - Anti-Crk p38 antibody [EP242Y] (ab45136)
  • Western blot - Anti-Crk p38 antibody [EP242Y] (ab45136)
  • Immunocytochemistry/ Immunofluorescence - Anti-Crk p38 antibody [EP242Y] (ab45136)

Key features and details

  • Rabbit monoclonal [EP242Y] to Crk p38
  • Suitable for: WB, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG

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Overview

  • Product name

    Anti-Crk p38 antibody [EP242Y]
    See all Crk p38 primary antibodies
  • Description

    Rabbit monoclonal [EP242Y] to Crk p38
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IFmore details
    Unsuitable for: Flow Cyt or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Crk p38 aa 250-350 (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: HeLa and HepG2 cell lysates. ICC/IF: HeLa cells.
  • General notes

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

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 99% Tissue culture supernatant, 0.1% BSA
  • Concentration information loading...
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EP242Y
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • MAPK Pathway
    • Signal Transduction
    • Adapters
    • Cytoplasmic
    • Cancer
    • Signal transduction
    • Other

Associated products

  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human CRK (Crk p38) knockout HeLa cell line (ab265097)
  • KO cell lysates

    • Human CRK (Crk p38) knockout HeLa cell lysate (ab257899)
  • Recombinant Protein

    • Recombinant Human Crk p38 protein (ab101046)

Applications

Our Abpromise guarantee covers the use of ab45136 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
ICC/IF
  • Application notes
    ICC/IF: 1/250 - 1/500.
    IP: 1/50.
    WB: 1/10000. Detects a band of approximately 37 kDa (predicted molecular weight: 34 kDa).

    Is unsuitable for Flow Cyt or IHC.


    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • Function

      The Crk-I and Crk-II forms differ in their biological activities. Crk-II has less transforming activity than Crk-I. Crk-II mediates attachment-induced MAPK8 activation, membrane ruffling and cell motility in a Rac-dependent manner. Involved in phagocytosis of apoptotic cells and cell motility via its interaction with DOCK1 and DOCK4.
    • Sequence similarities

      Belongs to the CRK family.
      Contains 1 SH2 domain.
      Contains 2 SH3 domains.
    • Domain

      The C-terminal SH3 domain function as a negative modulator for transformation and the N-terminal SH3 domain appears to function as a positive regulator for transformation.
      The SH2 domain mediates interaction with SHB.
    • Post-translational
      modifications

      Phosphorylation of Crk-II (40 kDa) gives rise to a 42 kDa form.
      Phosphorylated on Tyr-221 upon cell adhesion. Results in the negative regulation of the association with SH2- and SH3-binding partners, possibly by the formation of an intramolecular interaction of phosphorylated Tyr-221 with the SH2 domain. This leads finally to the down-regulation of the Crk signaling pathway.
    • Cellular localization

      Cytoplasm. Cell membrane. Translocated to the plasma membrane upon cell adhesion.
    • Target information above from: UniProt accession P46108 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 1398 Human
      • Entrez Gene: 12928 Mouse
      • Entrez Gene: 54245 Rat
      • Omim: 164762 Human
      • SwissProt: P46108 Human
      • SwissProt: Q64010 Mouse
      • SwissProt: Q63768 Rat
      • Unigene: 638121 Human
      • Unigene: 280125 Mouse
      • Unigene: 96136 Rat
      see all
    • Alternative names

      • Adapter molecule crk antibody
      • Avian sarcoma virus CT10 (v crk) oncogene homolog antibody
      • CRK antibody
      • CRK isoform 2 antibody
      • CRK isoform II antibody
      • CRK_HUMAN antibody
      • CRKII antibody
      • FLJ38130 antibody
      • OTTHUMP00000115366 antibody
      • OTTHUMP00000198330 antibody
      • p38 antibody
      • Proto oncogene C crk antibody
      • Proto-oncogene C-crk antibody
      • v crk avian sarcoma virus CT10 oncogene homolog antibody
      • v crk sarcoma virus CT10 oncogene homolog antibody
      • v crk sarcoma virus CT10 oncogene homolog (avian) antibody
      see all

    Images

    • Western blot - Anti-Crk p38 antibody [EP242Y] (ab45136)
      Western blot - Anti-Crk p38 antibody [EP242Y] (ab45136)
      All lanes : Anti-Crk p38 antibody [EP242Y] (ab45136) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : CRK knockout HeLa cell lysate
      Lane 3 : HepG2 cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Predicted band size: 34 kDa
      Observed band size: 38 kDa
      why is the actual band size different from the predicted?



      Lanes 1-3: Merged signal (red and green). Green - ab45136 observed at 38 kDa. Red - loading control ab7291 observed at 50 kDa. 

       ab45136 Anti-Crk p38 antibody [EP242Y] was shown to specifically react with Crk in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265097 (knockout cell lysate ab257899) was used.  Wild-type and Crk knockout samples were subjected to SDS-PAGE.  ab45136 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively.  Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-Crk p38 antibody [EP242Y] (ab45136)
      Western blot - Anti-Crk p38 antibody [EP242Y] (ab45136)
      Anti-Crk p38 antibody [EP242Y] (ab45136) at 1/10000 dilution + HepG2 cell lysate at 10 µg

      Secondary
      Goat anti-rabbit HRP at 1/2000 dilution

      Predicted band size: 34 kDa
      Observed band size: 37 kDa why is the actual band size different from the predicted?

    • Immunocytochemistry/ Immunofluorescence - Anti-Crk p38 antibody [EP242Y] (ab45136)
      Immunocytochemistry/ Immunofluorescence - Anti-Crk p38 antibody [EP242Y] (ab45136)
      Immunofluorescent staining of HeLa cells using ab45136 at a 1/250 dilution.

    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

    Datasheets and documents

    • Datasheet
  • References (1)

    Publishing research using ab45136? Please let us know so that we can cite the reference in this datasheet.

    ab45136 has been referenced in 1 publication.

    • Ahn J  et al. The metastasis gene NEDD9 product acts through integrin ß3 and Src to promote mesenchymal motility and inhibit amoeboid motility. J Cell Sci 125:1814-26 (2012). WB . PubMed: 22328516

    Customer reviews and Q&As

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

    Western blot abreview for Anti-Crk p38 antibody [EP242Y]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (mouse brain)
    Loading amount
    10 µg
    Specification
    mouse brain
    Gel Running Conditions
    Reduced Denaturing (12% SDS-PAGE)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted May 18 2011

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