Product nameAnti-Notch1 antibody [mN1A]
See all Notch1 primary antibodies
DescriptionMouse monoclonal [mN1A] to Notch1
Specificityab128076 recognizes intracellular domain of Notch1 protein, mainly its activated form. The unprocessed Notch1 protein is recognized with lower affinity.
Tested applicationsSuitable for: WB, IP, IHC-Fr, ICC, Flow Cytmore details
Species reactivityReacts with: Mouse, Human
Does not react with: Rat
Fusion protein corresponding to Mouse Notch1. GST fusion protein containing cdc10-NCR region of mouse Notch1
Database link: 4851
Storage instructionsShipped 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 bufferpH: 7.40
Preservative: 0.1% Sodium azide
Constituent: 99% PBS
Concentration information loading...
PurityProtein A purified
Purification notesPurified from cell culture supernatant. Purity >95% by SDS-PAGE.
Our Abpromise guarantee covers the use of ab128076 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. Predicted molecular weight: 272 kDa.|
|IP||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration.|
|ICC||Use at an assay dependent concentration.|
|Flow Cyt||Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, 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 [mN1A] (ab128076) at 1/1000 dilution
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 - ab128076 observed at 110 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab128076 was shown to recognize NOTCH1 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% Milk. Ab128076 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Overlay histogram showing SH-SY5Y cells stained with ab128076 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab128076, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
This product has been referenced in:
- Lin Y et al. Isoliquiritigenin inhibits the proliferation and induces the differentiation of human glioma stem cells. Oncol Rep 39:687-694 (2018). Read more (PubMed: 29251326) »
- Wang P et al. Basic fibroblast growth factor reduces scar by inhibiting the differentiation of epidermal stem cells to myofibroblasts via the Notch1/Jagged1 pathway. Stem Cell Res Ther 8:114 (2017). IF, IHC ; Rabbit . Read more (PubMed: 28511663) »