Product nameAnti-Notch1 antibody - ChIP Grade
See all Notch1 primary antibodies
DescriptionRabbit polyclonal to Notch1 - ChIP Grade
Tested applicationsSuitable for: WB, IHC-FoFr, ChIP, ICC/IF, IHC-Fr, IHC-Pmore details
Species reactivityReacts with: Mouse, Human
Predicted to work with: Rat
Synthetic peptide within Human Notch1 aa 2400-2500 (C terminal). The exact sequence is proprietary.
Database link: P46531
- Breast carcinoma, ovarian carcinoma.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.6
Preservative: 0.1% Sodium azide
Constituents: 0.0268% PBS, 1% BSA
Concentration information loading...
PurityImmunogen affinity purified
ChIP Related Products
Our Abpromise guarantee covers the use of ab27526 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration. PubMed: 18156632|
|IHC-FoFr||Use at an assay dependent concentration.|
|ChIP||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration.|
|IHC-P||1/200. for 10 minutes at room temperature. Staining of Formalin-fixed tissues requires boiling tissue sections in 10mM citrate buffer, pH 6.0 for 10 minutes followed by cooling at room temperature for 20 minutes.|
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
ChIP analysis using ab27526 binding Notch1 in mouse pancreatic cancer primary cells. Cells were cross-linked for 12 minutes with 1% formaldehyde. Samples were incubated with the primary antibody at a dilution of 1/50 for 8 hours at 4°C. Protein binding was detected using real-time PCR.
Pol II as was used as a positive control and IgG as background control. GAPDH promoter was used as the non-binding control.
ChIP Dilution Buffer:
1.1% Triton X-100,
16.7mM Tris-HCl, pH 8.1,
ab27526 staining Notch 1 in Mouse pancreatic cancer primary cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with acetone/methanol (1:1) and blocked with 10% serum for 1 hour at room temperature. Samples were incubated with primary antibody (1/100 in PBS) for 4 hours. An Alexa Fluor®488-conjugated Donkey anti-rabbit IgG polyclonal (ab150073) (1/1000) was used as the secondary antibody. Nuclei were counterstained with DAPI.
Anti-Notch1 antibody - ChIP Grade (ab27526) at 1/500 dilution (in TBST for 8 hours at 4°C) + Whole cell lysate of human MiaPaCa cells at 50 µg
An HRP-conjugated Donkey anti-rabbit IgG polyclonal at 1/5000 dilution
Developed using the ECL technique.
Performed under non-reducing conditions.
Observed band size: 120,250 kDa why is the actual band size different from the predicted?
Additional bands at: 75 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 10 minutes
Blocking Step: 5% Milk for 1 hour at room temperature.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma labelling Notch1 with ab27526.
ab27526 staining Notch1 in mouse neural tube by immunohistochemistry (Frozen). Sections were paraformaldhyde fixed prior to blocking in 20% serum for 16 hours at 25°C and then incubated with ab27526 for 16 hours at 25°C. A Cy3 conjugated donkey polyclonal to rabbit Ig was used as the secondary antibody. Blue staining is nuclear staining with DAPI and the specific Notch signal is red.
ab27526 staining Notch 1 in Human pancreatic neoplastic lesion tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 10% serum for 1 hour at room temperature; antigen retrieval was by heat mediation in antigen unmasking solution. Samples were incubated with primary antibody (1/100 in PBS) for 8 hours at 4°C. A Biotin-conjugated Goat anti-rabbit IgG polyclonal (ab6720) (1/1000) was used as the secondary antibody.
ab27526 staining Notch1 in Mouse brain tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with paraformaldehyde and blocked with 3% serum for 30 minutes at 25°C. The sample was incubated with primary antibody (1/50 in 3% Horse serum - ab7484) at 4°C for 15 hours. An undiluted HRP-conjugated Horse anti-rabbit IgG polyclonal was used as the secondary antibody.
This product has been referenced in:
- Park JC et al. Nerve Growth Factor Stimulates Glioblastoma Proliferation through Notch1 Receptor Signaling. J Korean Neurosurg Soc 61:441-449 (2018). Read more (PubMed: 29991107) »
- Natsuizaka M et al. Interplay between Notch1 and Notch3 promotes EMT and tumor initiation in squamous cell carcinoma. Nat Commun 8:1758 (2017). Read more (PubMed: 29170450) »