Product nameAnti-Notch1 antibody
See all Notch1 primary antibodies
DescriptionRabbit polyclonal to Notch1
Tested applicationsSuitable for: IHC-Fr, IP, WB, Flow Cytmore details
Species reactivityReacts with: Human
- This antibody gave a positive signal in the following whole cell lysates: HEK293, SK N SH and Jurkat.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab65297 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use a concentration of 5 µg/ml.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 120 kDa (predicted molecular weight: 272 kDa).|
|Flow Cyt||Use at an assay dependent concentration. PubMed: 24136221
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
FunctionFunctions 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.
Tissue specificityIn fetal tissues most abundant in spleen, brain stem and lung. Also present in most adult tissues where it is found mainly in lymphoid tissues.
Involvement in diseaseDefects 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.
Sequence similaritiesBelongs to the NOTCH family.
Contains 5 ANK repeats.
Contains 36 EGF-like domains.
Contains 3 LNR (Lin/Notch) repeats.
modificationsSynthesized 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.
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.
Cellular localizationCell membrane and Nucleus. Following proteolytical processing NICD is translocated to the nucleus.
- Information by UniProt
- 9930111A19Rik antibody
- AOS5 antibody
- AOVD1 antibody
All lanes : Anti-Notch1 antibody (ab65297) at 1 µg/ml
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : NOTCH1 knockout HAP1 whole cell lysate
Lysates/proteins at 40 µg per lane.
Predicted band size: 272 kDa
Lanes 1 - 2: Merged signal (red and green). Green - ab65297 observed at 110 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab65297 was shown to recognize in wild-type HAP1 cells as signal was lost at the expected MW in NOTCH1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and NOTCH1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% NF Milk. ab65297 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml 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.
All lanes : ab65927 at 1 mg/ml
Lane 1 : Human HEK293 whole cell lysate
Lane 2 : Human SKNSH whole cell lysate
Lane 3 : Human Jurkat whole cell lysate
Lysates/proteins at 10 µg per lane.
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 272 kDa
Observed band size: 112 kDa why is the actual band size different from the predicted?
Exposure time: 8 minutes
Notch1 was immunoprecipitated using 0.5mg Hek293 whole cell extract, 5µg of Rabbit polyclonal to Notch1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hek293 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab65297.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 120kDa; Notch1
ab65297 has been referenced in 10 publications.
- Xue F et al. Neferine inhibits growth and migration of gastrointestinal stromal tumor cell line GIST-T1 by up-regulation of miR-449a. Biomed Pharmacother 109:1951-1959 (2019). PubMed: 30551450
- Zhang Y et al. Ailanthone up-regulates miR-449a to restrain acute myeloid leukemia cells growth, migration and invasion. Exp Mol Pathol 108:114-120 (2019). PubMed: 31002772
- Guarnieri AL et al. The miR-106b-25 cluster mediates breast tumor initiation through activation of NOTCH1 via direct repression of NEDD4L. Oncogene 37:3879-3893 (2018). PubMed: 29662198
- Zhao W & Dong L Long non-coding RNA HOTAIR overexpression improves premature ovarian failure by upregulating Notch-1 expression. Exp Ther Med 16:4791-4795 (2018). PubMed: 30542434
- Ma B et al. Cartilage oligomeric matrix protein is a novel notch ligand driving embryonic stem cell differentiation towards the smooth muscle lineage. J Mol Cell Cardiol 121:69-80 (2018). PubMed: 29981303
- Zhou W et al. BLOS2 negatively regulates Notch signaling during neural and hematopoietic stem and progenitor cell development. Elife 5:N/A (2016). WB ; Mouse . PubMed: 27719760
- Zhang P et al. G protein-coupled receptor 183 facilitates endothelial-to-hematopoietic transition via Notch1 inhibition. Cell Res 25:1093-107 (2015). PubMed: 26358189
- Yao M et al. The Notch pathway mediates the angiotensin II-induced synthesis of extracellular matrix components in podocytes. Int J Mol Med 36:294-300 (2015). PubMed: 25902289
- Hu GH et al. Delta-like ligand 4 (Dll4) predicts the prognosis of clear cell renal cell carcinoma, and anti-Dll4 suppresses tumor growth in vivo. Int J Clin Exp Pathol 7:2143-52 (2014). WB ; Human . PubMed: 24966922
- Liu H et al. Single-cell clones of liver cancer stem cells have the potential of differentiating into different types of tumor cells. Cell Death Dis 4:e857 (2013). Flow Cyt ; Human . PubMed: 24136221