Recombinant Anti-CBL antibody [YE323] - C-terminal (ab32027)
![Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)](https://www.abcam.com/ps/products/32/ab32027/Images/ab32027-411463-anti-cbl-antibody-ye323-c-terminal-westernblot-human-mouse-rat.jpg "Western blot - 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
Related conjugates and formulations
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
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
- Signaling Pathway
- Nuclear Signaling
- Nuclear Hormone Receptors
- Co-activators/co-repressors
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
KO cell lines
-
KO cell lysates
-
Positive Controls
-
Recombinant Protein
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 |
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
modificationsPhosphorylated 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
- 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
see all -
Alternative names
- 4732447J05Rik antibody
- C CBL antibody
- Cas Br M (murine) ecotropic retroviral transforming sequence antibody
see all
Images
-
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)](https://www.abcam.com/ps/products/32/ab32027/Images/ab32027-410226-anti-cbl-antibody-ye323-c-terminal-western-blot-human-knockout-lysate.jpg)
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)](https://www.abcam.com/ps/products/32/ab32027/Images/ab32027-6-anti-cbl-antibody-ye323-c-terminal-immunocytochemistry-jurkat-human.jpg)
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)](https://www.abcam.com/ps/products/32/ab32027/Images/ab32027-7-anti-cbl-antibody-ye323-c-terminal-flowcytometry-jurkat-human.jpg)
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)](https://www.abcam.com/ps/products/32/ab32027/Images/ab32027-299907-anti-cbl-antibody-ye323-c-terminal-western-blot.jpg)
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)](https://www.abcam.com/ps/products/32/ab32027/Images/ab32027-397395-anti-cbl-antibody-ye323-c-terminal-western-blot-wehi-2311.jpg)
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)](https://www.abcam.com/ps/products/32/ab32027/Images/ab32027-9-benefits-of-recombinant-antibodies.png)
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.
Datasheets and documents
-
SDS download
-
Datasheet download
References (11)
ab32027 has been referenced in 11 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
- Lu X et al. CTEN Inhibits Tumor Angiogenesis and Growth by Targeting VEGFA Through Down-Regulation of β-Catenin in Breast Cancer. Technol Cancer Res Treat 20:15330338211045506 (2021). PubMed: 34817293
- 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