重组Anti-CBL抗体[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
概述
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产品名称
Anti-CBL抗体[YE323] - C-terminal
参阅全部 CBL 一抗 -
描述
兔单克隆抗体[YE323] to CBL - C-terminal -
宿主
Rabbit -
经测试应用
适用于: WB, Flow Cyt (Intra), ICC/IFmore details
不适用于: IP -
种属反应性
与反应: Mouse, Rat, Human
预测可用于: Chicken -
免疫原
Synthetic peptide within Human CBL aa 850 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: P22681 -
表位
ab32027 reacts with an epitope located in the C terminal region of CBL. -
阳性对照
- 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.
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常规说明
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.
性能
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形式
Liquid -
存放说明
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. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
YE323 -
同种型
IgG -
研究领域
- 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
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab32027于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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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 |
说明 |
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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 |
靶标
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功能
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. -
通路
Protein modification; protein ubiquitination. -
疾病相关
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. -
序列相似性
Contains 1 Cbl-PTB (Cbl-type phosphotyrosine-binding) domain.
Contains 1 RING-type zinc finger.
Contains 1 UBA domain. -
结构域
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. -
翻译后修饰
Phosphorylated on tyrosine residues by EGFR, SYK, FYN and ZAP70 (By similarity). Phosphorylated on tyrosine residues by INSR. -
细胞定位
Cytoplasm. - Information by UniProt
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数据库链接
- 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 -
别名
- 4732447J05Rik antibody
- C CBL antibody
- Cas Br M (murine) ecotropic retroviral transforming sequence antibody
see all
图片
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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? -
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.
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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.
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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).
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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.
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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
实验方案
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.
数据表及文件
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SDS download
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Datasheet download
文献 (11)
ab32027 被引用在 11 文献中.
- 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