Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-CBL antibody [YE323] - BSA and Azide free (ab236075)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [YE323] to CBL - BSA and Azide free
  • Suitable for: ICC, Flow Cyt, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-CBL antibody [YE323] - BSA and Azide free
    See all CBL primary antibodies

  • Description

    Rabbit monoclonal [YE323] to CBL - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC, Flow Cyt, WBmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Chicken

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Epitope

    ab236075 reacts with an epitope located in the C terminal region of CBL.

  • Positive control

    • WB: HEK293T, HAP1, Jurkat, THP-1, WEHI-231, F9 and Raji cell lysates; Mouse thymus tissue lysate, Rat testis lysate, Rat thymus lysate. ICC: Jurkat cells. Flow Cyt: Jurkat cells.

  • General notes

    ab236075 is the carrier-free version of ab32027 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

    Ab236075 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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

Images

  • Western blot - Anti-CBL antibody [YE323] - BSA and Azide free (ab236075)
    Western blot - Anti-CBL antibody [YE323] - BSA and Azide free (ab236075)
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

    Lane 1 : Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate at 15 µg
    Lane 2 : Rat testis lysate at 20 µg
    Lane 3 : Rat thymus lysate at 20 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/20000 dilution

    Predicted band size: 99 kDa
    Observed band size: 110 kDa
    why is the actual band size different from the predicted?



    This data was developed using ab32027, the same antibody clone in a different buffer formulation.

  • Western blot - Anti-CBL antibody [YE323] - BSA and Azide free (ab236075)
    Western blot - Anti-CBL antibody [YE323] - BSA and Azide free (ab236075)
    All lanes : Anti-CBL antibody [YE323] - C-terminal (ab32027) at 1/1000 dilution (unpurified)

    Lane 1 : Wild-type HEK293T cell lysate
    Lane 2 : CBL knockout HEK293T cell lysate
    Lane 3 : THP-1 cell lysate
    Lane 4 : Raji 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: 99 kDa
    Observed band size: 110 kDa why is the actual band size different from the predicted?



    This data was developed using ab32027, the same antibody clone in a different buffer formulation.

    Lanes 1-4: Merged signal (red and green). Green - ab32027 observed at 110 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab32027 Anti-CBL antibody [YE323] - C-terminal was shown to specifically react with CBL in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267245 (knockout cell lysate ab257200) was used. Wild-type and CBL knockout samples were subjected to SDS-PAGE. ab32027 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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-CBL antibody [YE323] - BSA and Azide free (ab236075)
    Immunocytochemistry - Anti-CBL antibody [YE323] - BSA and Azide free (ab236075)
    This data was developed using ab32027, the same antibody clone in a different buffer formulation.
    Immunocytochemistry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling CBL with purified ab32027 at 1/50 dilution (4.26 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-CBL antibody [YE323] - BSA and Azide free (ab236075)
    Flow Cytometry - Anti-CBL antibody [YE323] - BSA and Azide free (ab236075)
    This data was developed using ab32027, the same antibody clone in a different buffer formulation.
    Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling CBL with purified ab32027 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-CBL antibody [YE323] - BSA and Azide free (ab236075)
    Western blot - Anti-CBL antibody [YE323] - BSA and Azide free (ab236075)

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: CBL knockout HAP1 whole cell lysate (20 µg)
    Lane 3: THP1 whole cell lysate (20 µg)
    Lane 4: Raji whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab32027 observed at 100 kDa. Red - loading control, ab9484, observed at 37 kDa.

    Unpurified ab32027 was shown to specifically react with CBL in wild-type HAP1 cells as signal was lost in CBL knockout cells. Wild-type and CBL knockout samples were subjected to SDS-PAGE. ab32027 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32027).

  • Western blot - Anti-CBL antibody [YE323] - BSA and Azide free (ab236075)
    Western blot - Anti-CBL antibody [YE323] - BSA and Azide free (ab236075)
    All lanes : Anti-CBL antibody [YE323] - C-terminal (ab32027) at 1/1000 dilution

    Lane 1 : WEHI-231 (Mouse B cell lymphoma B lymphocyte) cell lysate
    Lane 2 : F9 (Mouse embryonal carcinoma epithelial cell) cell lysate
    Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) cell lysate
    Lane 4 : Mouse thymus lysate

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 99 kDa
    Observed band size: 110 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer and concentration: 5% NFDM/TBST

     

     Exposure time: 30 seconds

  • Anti-CBL antibody [YE323] - BSA and Azide free (ab236075)
    Anti-CBL antibody [YE323] - BSA and Azide free (ab236075)

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