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    products/primary-antibodies/cbl-antibody-ye323-c-terminal-ab32027.pdf

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Cell Biology Proteolysis / Ubiquitin Proteasome / Ubiquitin Ubiquitin E3 Enzymes RING Finger E3 Ligase
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Validated using a knockout cell lineRecombinantRabMAb

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

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Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)
  • Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)
  • Immunocytochemistry/ Immunofluorescence - Anti-CBL antibody [YE323] - C-terminal (ab32027)
  • Flow Cytometry (Intracellular) - Anti-CBL antibody [YE323] - C-terminal (ab32027)
  • Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)
  • Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)
  • 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: WB, Flow Cyt (Intra), ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 647 Carrier Free PE

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Human CBL knockout HEK-293T cell line (ab267245)

View more associated products

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

    Suitable for: WB, Flow Cyt (Intra), ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Chicken
  • 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/IF: Jurkat cells. Flow Cyt (intra): 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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    YE323
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteasome / Ubiquitin
    • Ubiquitin E3 Enzymes
    • RING Finger E3 Ligase
    • Signal Transduction
    • Adapters
    • Cytoplasmic
    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • Nuclear Hormone Receptors
    • Co-activators/co-repressors
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Cancer susceptibility
    • Proto-oncogenes
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • Nuclear Receptors
    • Co-activators/co-repressors
    • Cancer
    • Signal transduction
    • Nuclear signaling
    • Nuclear hormone receptors
    • Coactivators and repressors
    • Cancer
    • Oncoproteins/suppressors
    • Oncoproteins
    • Other

Associated products

  • Alternative Versions

    • Alexa Fluor® 647 Anti-CBL antibody [YE323] - C-terminal (ab224916)
    • PE Anti-CBL antibody [YE323] - C-terminal (ab224917)
    • Anti-CBL antibody [YE323] - BSA and Azide free (ab236075)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human CBL knockout HEK-293T cell line (ab267245)
  • KO cell lysates

    • Human CBL knockout HEK-293T cell lysate (ab257200)
  • Positive Controls

    • Jurkat whole cell lysate (ab7899)
  • Recombinant Protein

    • Recombinant Human CBL protein (ab114527)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32027 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
1/1000. Detects a band of approximately 120 kDa (predicted molecular weight: 99 kDa).

For unpurified use at 1/5000

Flow Cyt (Intra)
1/30.

For unpurified use at 1/100  ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF
1/50.

For unpurified use at 1/100

Notes
WB
1/1000. Detects a band of approximately 120 kDa (predicted molecular weight: 99 kDa).

For unpurified use at 1/5000

Flow Cyt (Intra)
1/30.

For unpurified use at 1/100  ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF
1/50.

For unpurified use at 1/100

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.
  • Target information above from: UniProt accession P22681 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 374038 Chicken
    • Entrez Gene: 867 Human
    • Entrez Gene: 12402 Mouse
    • Entrez Gene: 500985 Rat
    • Omim: 165360 Human
    • SwissProt: P22681 Human
    • SwissProt: P22682 Mouse
    • Unigene: 504096 Human
    • Unigene: 266871 Mouse
    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] - 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 : 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?

  • 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/ Immunofluorescence - Anti-CBL antibody [YE323] - C-terminal (ab32027)
    Immunocytochemistry/ Immunofluorescence - 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 (Intracellular) - Anti-CBL antibody [YE323] - C-terminal (ab32027)
    Flow Cytometry (Intracellular) - Anti-CBL antibody [YE323] - C-terminal (ab32027)

    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] - 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)

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

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (10)

Publishing research using ab32027? Please let us know so that we can cite the reference in this datasheet.

ab32027 has been referenced in 10 publications.

  • Lei X  et al. circTLK1 facilitates the proliferation and metastasis of renal cell carcinoma by regulating miR-495-3p/CBL axis. Open Life Sci 16:362-374 (2021). PubMed: 33954256
  • Xie G  et al. Conjugated secondary 12a-hydroxylated bile acids promote liver fibrogenesis. EBioMedicine 66:103290 (2021). PubMed: 33752128
  • Zhang D  et al. MicroRNA-935 Directly Targets FZD6 to Inhibit the Proliferation of Human Glioblastoma and Correlate to Glioma Malignancy and Prognosis. Front Oncol 11:566492 (2021). PubMed: 33791198
  • Xia Z  et al. miR-652 Promotes Proliferation and Migration of Uveal Melanoma Cells by Targeting HOXA9. Med Sci Monit 25:8722-8732 (2019). PubMed: 31740654
  • Zhao T  et al. DNA methylation-regulated QPCT promotes sunitinib resistance by increasing HRAS stability in renal cell carcinoma. Theranostics 9:6175-6190 (2019). PubMed: 31534544
  • Duong MN  et al. Chronic TCR-MHC (self)-interactions limit the functional potential of TCR affinity-increased CD8 T lymphocytes. J Immunother Cancer 7:284 (2019). PubMed: 31690351
  • Schmid FM  et al. IFT20 modulates ciliary PDGFRa signaling by regulating the stability of Cbl E3 ubiquitin ligases. J Cell Biol 217:151-161 (2018). PubMed: 29237719
  • Peng L  et al. Huntingtin-Interacting Protein 1-Related Protein Plays a Critical Role in Dendritic Development and Excitatory Synapse Formation in Hippocampal Neurons. Front Mol Neurosci 10:186 (2017). WB, ICC ; Rat . PubMed: 28663723
  • Jiang R  et al. The long noncoding RNA lnc-EGFR stimulates T-regulatory cells differentiation thus promoting hepatocellular carcinoma immune evasion. Nat Commun 8:15129 (2017). WB, IP . PubMed: 28541302
  • Walsh AM & Lazzara MJ Regulation of EGFR trafficking and cell signaling by Sprouty2 and MIG6 in lung cancer cells. J Cell Sci 126:4339-48 (2013). WB ; Human . PubMed: 23868981

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