重组Anti-Notch1抗体[EP1238Y] (ab52627)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1238Y] to Notch1
- Suitable for: IHC-P, Flow Cyt (Intra), WB, ICC/IF
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
概述
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产品名称
Anti-Notch1抗体[EP1238Y]
参阅全部 Notch1 一抗 -
描述
兔单克隆抗体[EP1238Y] to Notch1 -
宿主
Rabbit -
特异性
80% identities with Notch 2 and 81% with Notch 3 -
经测试应用
适用于: IHC-P, Flow Cyt (Intra), WB, ICC/IFmore details
不适用于: IP -
种属反应性
与反应: Mouse, Human
预测可用于: Cow -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HeLa, HAP1, MOLT-4, HEK-293 cell lysate. Mouse brain lysate. IHC-P: Human brain tissue. ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cells.
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常规说明
Notch 1 (encoded by the Notch 1 gene) is a member of the Notch family of receptors. When bound to a ligand, the Notch1 receptor sends signals critical for normal development of many tissues throughout the body. Notch1 signalling supports the determination and specialization of cells as well as playing a role in cell proliferation, differentiation, and apoptosis.
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. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EP1238Y -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Alexa Fluor® 647 Anti-Notch1 antibody [EP1238Y] (ab194122)
- Anti-Notch1 antibody [EP1238Y] - BSA and Azide free (ab221603)
- Anti-Notch1 antibody [EP1238Y] - Low endotoxin, Azide free (ab246693)
- Alexa Fluor® 555 Anti-Notch1 antibody [EP1238Y] (ab315420)
- Alexa Fluor® 750 Anti-Notch1 antibody [EP1238Y] (ab321656)
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab52627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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IHC-P | (9) |
1/100 - 1/150. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/130 - 1/200.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (7) |
1/1000 - 1/2000. Detects a band of approximately 125 kDa.
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ICC/IF |
1/100 - 1/150.
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说明 |
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IHC-P
1/100 - 1/150. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/130 - 1/200. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/1000 - 1/2000. Detects a band of approximately 125 kDa. |
ICC/IF
1/100 - 1/150. |
靶标
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功能
Functions as a receptor for membrane-bound ligands Jagged1, Jagged2 and Delta1 to regulate cell-fate determination. Upon ligand activation through the released notch intracellular domain (NICD) it forms a transcriptional activator complex with RBPJ/RBPSUH and activates genes of the enhancer of split locus. Affects the implementation of differentiation, proliferation and apoptotic programs. May be important for normal lymphocyte function. In altered form, may contribute to transformation or progression in some T-cell neoplasms. Involved in the maturation of both CD4+ and CD8+ cells in the thymus. May be important for follicular differentiation and possibly cell fate selection within the follicle. During cerebellar development, may function as a receptor for neuronal DNER and may be involved in the differentiation of Bergmann glia. -
组织特异性
In fetal tissues most abundant in spleen, brain stem and lung. Also present in most adult tissues where it is found mainly in lymphoid tissues. -
疾病相关
Defects in NOTCH1 are a cause of bicuspid aortic valve (BAV) [MIM:109730]. A common defect in the aortic valve in which two rather than three leaflets are present. It is often associated with aortic valve calcification and insufficiency. In extreme cases, the blood flow may be so restricted that the left ventricle fails to grow, resulting in hypoplastic left heart syndrome. -
序列相似性
Belongs to the NOTCH family.
Contains 5 ANK repeats.
Contains 36 EGF-like domains.
Contains 3 LNR (Lin/Notch) repeats. -
翻译后修饰
Synthesized in the endoplasmic reticulum as an inactive form which is proteolytically cleaved by a furin-like convertase in the trans-Golgi network before it reaches the plasma membrane to yield an active, ligand-accessible form. Cleavage results in a C-terminal fragment N(TM) and a N-terminal fragment N(EC). Following ligand binding, it is cleaved by TNF-alpha converting enzyme (TACE) to yield a membrane-associated intermediate fragment called notch extracellular truncation (NEXT). This fragment is then cleaved by presenilin dependent gamma-secretase to release a notch-derived peptide containing the intracellular domain (NICD) from the membrane.
Phosphorylated.
O-glycosylated on the EGF-like domains. Contains both O-linked fucose and O-linked glucose.
Ubiquitinated; undergoes 'Lys-29'-linked polyubiquitination catalyzed by ITCH. -
细胞定位
Cell membrane and Nucleus. Following proteolytical processing NICD is translocated to the nucleus. - Information by UniProt
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数据库链接
- Entrez Gene: 767866 Cow
- Entrez Gene: 4851 Human
- Entrez Gene: 18128 Mouse
- Omim: 190198 Human
- SwissProt: P46531 Human
- SwissProt: Q01705 Mouse
- Unigene: 495473 Human
- Unigene: 290610 Mouse
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别名
- 9930111A19Rik antibody
- AOS5 antibody
- AOVD1 antibody
see all
图片
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All lanes : Anti-Notch1 antibody [EP1238Y] (ab52627) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : NOTCH1 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.Lanes 1- 2: Merged signal (red and green). Green - ab52627 observed at 110 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab52627 was shown to react with Notch1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261762 (knockout cell lysate ab257006) was used. Wild-type HeLa and NOTCH1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab52627 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 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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Immunofluorescent staining of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells fixed with 4% PFA using purified ab52627 at a dilution of 1/150.
An Alexa Fluor® 555 goat anti-rabbit was used as the secondary and the sample was stained with DAPI. An Alexa Fluor® 555 goat anti-rabbit was used at a dilution of 1/500 as the negative control and is shown in the bottom right hand panel.
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All lanes : Anti-Notch1 antibody [EP1238Y] (ab52627) at 1/1000 dilution
Lane 1 : Wild-type HL-60 cell lysate
Lane 2 : NOTCH1 knockout HL-60 cell lysate
Lane 3 : Wild-type HAP1 cell lysate
Lane 4 : NOTCH1 knockout HAP1 cell lysate
Lane 5 : MOLT-4 cell lysate
Lane 6 : HepG2 cell lysate
Lysates/proteins at 20 µg per lane.
Developed using the ECL technique.
Performed under reducing conditions.
Observed band size: 100 kDa why is the actual band size different from the predicted?Western blot: Anti-NOTCH1 antibody [EP1238Y] (ab52627) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab52627 was shown to bind specifically to NOTCH1. A band was observed at 100 kDa in wild-type HL-60 cell lysates with no signal observed at this size in NOTCH1 knockout cell line. To generate this image, wild-type and NOTCH1 knockout HL-60 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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All lanes : Anti-Notch1 antibody [EP1238Y] (ab52627) at 1 µg/ml
Lane 1 : Wild-type HAP1 cell lysate at 40 µg
Lane 2 : NOTCH1 knockout HAP1 cell lysate at 40 µg
Lane 3 : MOLT-4 cell lysate at 20 µg
Lane 4 : HepG2 cell lysate at 20 µg
Performed under reducing conditions.
Observed band size: 105 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab52627 observed at 105 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab52627 was shown to react with Notch1 in wild-type HAP1 cells in western blot. Loss of signal was observed when NOTCH1 knockout sample was used. Wild-type and NOTCH1 knockout HAP1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk before incubation with ab52627 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 hour at room temperature before imaging.
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Anti-Notch1 antibody [EP1238Y] (ab52627) at 1/2000 dilution (Purified) + Mouse brain at 10 µg
Secondary
HRP goat anti-rabbit (H+L) at 1/1000 dilution
Observed band size: 125 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
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Anti-Notch1 antibody [EP1238Y] (ab52627) at 1/2000 dilution (Purified) + HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate at 10 µg
Secondary
HRP goat anti-rabbit (H+L) at 1/1000 dilution
Observed band size: 125 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
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Verification of gene expression array data by immunohistochemical analysis of Notch 1 expression in subcutaneous tumors and lung metastases from a human melanoma (MeWo) xenograft experiment in mice.
Immunohistochemical staining for Notch1 expression (ab52627, red) in subcutaneous tumors and lung metastases (both panels) of MeWo (Human malignant melanoma cell line) cells.
All scale bars: 50 μm.
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Immunohistochemical staining of paraffin-embedded human brain with purified ab52627 at a dilution of 1/150.
A prediluted HRP polymer for rabbit IgG was used as the secondary and the sample was stained with hematoxylin. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemical staining of paraffin-embedded human brain with unpurified ab52627 at a dilution of 1/100.
A prediluted HRP polymer for rabbit IgG was used as the secondary and the sample was stained with hematoxylin. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunofluorescent staining of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells fixed with 4% PFA using unpurified ab52627 at a dilution of 1/100.
An Alexa Fluor® 555 goat anti-rabbit was used as the secondary and the sample was stained with DAPI. An Alexa Fluor® 555 goat anti-rabbit was used at a dilution of 1/500 as the negative control and is shown in the bottom right hand panel.
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Intracellular Flow Cytometry analysis of permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells (2% PFA, pink) withpurified ab52627 at a 1/200 dilution, or negative control rabbit monoclonal IgG (green). The secondary antibody was FITC goat anti-rabbit.
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 (201)
ab52627 被引用在 201 文献中.
- Zhang W et al. Single-cell sequencing reveals SATB2/NOTCH1 signaling promotes the progression of malignancy of epithelial cells from papillary thyroid cancer. Mol Carcinog 63:22-33 (2024). PubMed: 37877736
- Mukherjee D et al. Fetal pulmonary hypertension: dysregulated microRNA-34c-Notch1 axis contributes to impaired angiogenesis in an ovine model. Pediatr Res 93:551-558 (2023). PubMed: 35717485
- Hu HJ et al. Effects of hyperbaric oxygen on Notch signaling pathway after severe carbon monoxide poisoning in mice. Med Gas Res 13:23-28 (2023). PubMed: 35946219
- Pei J et al. Epigenetic regulator KDM4A activates Notch1-NICD-dependent signaling to drive tumorigenesis and metastasis in breast cancer. Transl Oncol 28:101615 (2023). PubMed: 36592610
- Gu NX et al. Frizzled 7 modulates goblet and Paneth cell fate, and maintains homeostasis in mouse intestine. Development 150:N/A (2023). PubMed: 36691900