Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-CBL antibody [YE323] - C-terminal (ab32027)

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 - C-terminal
  • Suitable for: ICC, WB, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-CBL antibody [YE323] - C-terminal
    See all CBL primary antibodies

  • Description

    Rabbit monoclonal [YE323] to CBL - C-terminal

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC
    Human
    WB
    Mouse
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human CBL aa 850 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: P22681

  • Epitope

    ab32027 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

    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.

Properties

Images

  • Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)
    Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)
    Lane 1 : Anti-CBL antibody [YE323] - C-terminal (ab32027) at 1/1000 dilution (Purified)
    Lanes 2-3 : Anti-CBL antibody [YE323] - C-terminal (ab32027) 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
    Lane 1 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/2000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
    Lanes 2-3 : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/2000 dilution

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

  • Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)
    Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)
    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?



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

    Unpurified 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] - C-terminal (ab32027)
    Immunocytochemistry - Anti-CBL antibody [YE323] - C-terminal (ab32027)
    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] - C-terminal (ab32027)
    Flow Cytometry - Anti-CBL antibody [YE323] - C-terminal (ab32027)
    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] - C-terminal (ab32027)
    Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)

    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.

  • Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)
    Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)
    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] - C-terminal (ab32027)
    Anti-CBL antibody [YE323] - C-terminal (ab32027)

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