Validated using a knockout cell lineRabMAb

Anti-Crk p38 antibody [EP242Y] (ab45136)

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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: Human
  • Isotype: IgG

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Carrier Free

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: Human
    Predicted to work with: Mouse, Rat

  • 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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

Properties

Applications

The Abpromise guarantee

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 (1)
1/10000. Detects a band of approximately 37 kDa (predicted molecular weight: 34 kDa).
ICC/IF
1/250 - 1/500.
Notes
WB
1/10000. Detects a band of approximately 37 kDa (predicted molecular weight: 34 kDa).
ICC/IF
1/250 - 1/500.
Application notes
Is unsuitable for Flow Cyt or IP.

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.
  • Information by UniProt

  • Database links

    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

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

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

 Excellent
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
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Verified customer

Submitted May 18 2011

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