Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-CrkL antibody [Y244] - BSA and Azide free (ab247210)

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Western blot - Anti-CrkL antibody [Y244] - BSA and Azide free (ab247210)
  • Immunocytochemistry - Anti-CrkL antibody [Y244] - BSA and Azide free (ab247210)
  • Immunoprecipitation - Anti-CrkL antibody [Y244] - BSA and Azide free (ab247210)
  • Anti-CrkL antibody [Y244] - BSA and Azide free (ab247210)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y244] to CrkL - BSA and Azide free
  • Suitable for: ICC, WB, IHC-P, IP
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-CrkL antibody [Y244] - BSA and Azide free
    See all CrkL primary antibodies

  • Description

    Rabbit monoclonal [Y244] to CrkL - BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    This antibody recognises CrkL (Crk-like)protein.

  • Tested applications

    Suitable for: ICC, WB, IHC-P, IPmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

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

  • Positive control

    • WB: HeLa and K562 cell lysates. ICC: K-562 cells IP: K-562 whole cell lysate.

  • General notes

    ab247210 is the carrier-free version of ab32018 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.

    Ab247210 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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 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

Applications

Our Abpromise guarantee covers the use of ab247210 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
ICC Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 36 kDa (predicted molecular weight: 33 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP Use at an assay dependent concentration.

Target

  • Function

    May mediate the transduction of intracellular signals.

  • Sequence similarities

    Belongs to the CRK family.
    Contains 1 SH2 domain.
    Contains 2 SH3 domains.
  • Information by UniProt

  • Database links

    • Alternative names

      • Crk L antibody
      • Crk like protein antibody
      • Crk-like protein antibody
      • Crkl antibody
      • CRKL_HUMAN antibody
      • Crkol antibody
      • HGNC:2363 antibody
      • Oncogene CrkL antibody
      • V crk avian sarcoma virus CT10 oncogene homolog like antibody
      • v crk sarcoma virus CT10 oncogene homolog (avian) like antibody
      • V crk sarcoma virus CT10 oncogene homolog avian like antibody
      see all

    Images

    • Western blot - Anti-CrkL antibody [Y244] - BSA and Azide free (ab247210)
      Western blot - Anti-CrkL antibody [Y244] - BSA and Azide free (ab247210)
      All lanes : Anti-CrkL antibody [Y244] (ab32018) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : CRKL knockout HeLa cell lysate
      Lane 3 : K-562 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: 33 kDa
      Observed band size: 37 kDa
      why is the actual band size different from the predicted?



      This data was developed using the same antibody clone in a different buffer formulation (ab32018).

      Lanes 1-3: Merged signal (red and green). Green - ab32018 observed at 37 kDa. Red - loading control ab7291 observed at 50 kDa.

      ab32018 Anti-CrkL antibody [Y244] was shown to specifically react with CrkL in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265993 (knockout cell lysate ab257397) was used. Wild-type and CrkL knockout samples were subjected to SDS-PAGE. ab32018 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.

       

       

    • Immunocytochemistry - Anti-CrkL antibody [Y244] - BSA and Azide free (ab247210)
      Immunocytochemistry - Anti-CrkL antibody [Y244] - BSA and Azide free (ab247210)
      This data was developed using the same antibody clone in a different buffer formulation (ab32018).
      Immunocytochemistry analysis of K-562 (human chronic myelogenous leukemia lymphoblast) labeling CrkL with purified ab32018 at 1/100 dilution (10 µg/ml). Cells were fixed with 100% methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml) was used as counterstain. Nuclei were stained blue with DAPI.
      Negative control: PBS instead of the primary antibody.
    • Immunoprecipitation - Anti-CrkL antibody [Y244] - BSA and Azide free (ab247210)
      Immunoprecipitation - Anti-CrkL antibody [Y244] - BSA and Azide free (ab247210)

      This data was developed using ab32018, the same antibody clone in a different buffer formulation.
      Purified ab32018 at 1/50 dilution (2µg) immunoprecipitating CrkL in K-562 whole cell lysate.
      Lane 1 (input): K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate 10µg
      Lane 2 (+): ab32018 + K-562 whole cell lysate.
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32018 in K-562 whole cell lysate.
      VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
      Blocking Buffer and concentration: 5% NFDM/TBST.
      Diluting buffer and concentration: 5% NFDM/TBST.
      Observed band size: 37 kDa

    • Anti-CrkL antibody [Y244] - BSA and Azide free (ab247210)
      Anti-CrkL antibody [Y244] - BSA and Azide free (ab247210)

    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 ab247210? Please let us know so that we can cite the reference in this datasheet.

    ab247210 has not yet been referenced specifically in any publications.

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