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Validated using a knockout cell lineRecombinantRabMAb

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

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Western blot - Anti-CBL antibody [YE323] - BSA and Azide free (ab236075)
  • Western blot - Anti-CBL antibody [YE323] - BSA and Azide free (ab236075)
  • Immunocytochemistry/ Immunofluorescence - Anti-CBL antibody [YE323] - BSA and Azide free (ab236075)
  • Flow Cytometry (Intracellular) - Anti-CBL antibody [YE323] - BSA and Azide free (ab236075)
  • Western blot - Anti-CBL antibody [YE323] - BSA and Azide free (ab236075)
  • Western blot - Anti-CBL antibody [YE323] - BSA and Azide free (ab236075)
  • 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: Flow Cyt (Intra), WB, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 647 PE Unconjugated

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: Flow Cyt (Intra), WB, ICC/IFmore 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/IF: Jurkat cells. Flow Cyt (intra): Jurkat cells.

  • General notes

    ab236075 is the carrier-free version of ab32027.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    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.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. 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.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab236075 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
Flow Cyt (Intra)
Use at an assay dependent concentration.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB
Use at an assay dependent concentration. Detects a band of approximately 120 kDa (predicted molecular weight: 99 kDa).
ICC/IF
Use at an assay dependent concentration.
Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB
Use at an assay dependent concentration. Detects a band of approximately 120 kDa (predicted molecular weight: 99 kDa).
ICC/IF
Use at an assay dependent concentration.
Application notes
Is unsuitable for IP.

Target

  • Function

    Participates in signal transduction in hematopoietic cells. Adapter protein that functions as a negative regulator of many signaling pathways that start from receptors at the cell surface. Acts as an E3 ubiquitin-protein ligase, which accepts ubiquitin from specific E2 ubiquitin-conjugating enzymes, and then transfers it to substrates promoting their degradation by the proteasome. Recognizes activated receptor tyrosine kinases, including PDGFA, EGF and CSF1, and terminates signaling.

  • Pathway

    Protein modification; protein ubiquitination.

  • Involvement in disease

    Defects in CBL are the cause of Noonan syndrome-like disorder (NSL) [MIM:613563]. NSL is a syndrome characterized by a phenotype reminiscent of Noonan syndrome. Clinical features are highly variable, including facial dysmorphism, short neck, developmental delay, hyperextensible joints and thorax abnormalities with widely spaced nipples. The facial features consist of triangular face with hypertelorism, large low-set ears, ptosis, and flat nasal bridge. Some patients manifest cardiac defects.

  • Sequence similarities

    Contains 1 Cbl-PTB (Cbl-type phosphotyrosine-binding) domain.
    Contains 1 RING-type zinc finger.
    Contains 1 UBA domain.

  • Domain

    The RING-type zinc finger domain mediates binding to an E2 ubiquitin-conjugating enzyme.
    The N-terminus is composed of the phosphotyrosine binding (PTB) domain, a short linker region and the RING-type zinc finger. The PTB domain, which is also called TKB (tyrosine kinase binding) domain, is composed of three different subdomains: a four-helix bundle (4H), a calcium-binding EF hand and a divergent SH2 domain.

  • Post-translational
    modifications

    Phosphorylated on tyrosine residues by EGFR, SYK, FYN and ZAP70 (By similarity). Phosphorylated on tyrosine residues by INSR.

  • Cellular localization

    Cytoplasm.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • 4732447J05Rik antibody
    • C CBL antibody
    • Cas Br M (murine) ecotropic retroviral transforming sequence antibody
    • Casitas B lineage lymphoma proto oncogene antibody
    • Casitas B-lineage lymphoma proto-oncogene antibody
    • CBL 2 antibody
    • cbl antibody
    • CBL_HUMAN antibody
    • CBL2 antibody
    • E3 ubiquitin protein ligase CBL antibody
    • E3 ubiquitin-protein ligase CBL antibody
    • Oncogene CBL2 antibody
    • Proto oncogene c CBL antibody
    • Proto-oncogene c-CBL antibody
    • RGD1561386 antibody
    • RING finger protein 55 antibody
    • RNF55 antibody
    • Signal transduction protein CBL antibody
    see all

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 : 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
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/2000 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/ Immunofluorescence - Anti-CBL antibody [YE323] - BSA and Azide free (ab236075)
    Immunocytochemistry/ Immunofluorescence - 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 (Intracellular) - Anti-CBL antibody [YE323] - BSA and Azide free (ab236075)
    Flow Cytometry (Intracellular) - Anti-CBL antibody [YE323] - BSA and Azide free (ab236075)

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

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 ab236075? Please let us know so that we can cite the reference in this datasheet.

ab236075 has not yet been referenced specifically in any publications.

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