Anti-activated Notch1 antibody (ab83253)

Overview

  • Product nameAnti-activated Notch1 antibody
    See all activated Notch1 primary antibodies
  • Description
    Rabbit polyclonal to activated Notch1
  • Tested applicationsSuitable for: WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide derived from within residues 1750 - 1850 of Human activated Notch1.

    (Peptide available as ab93067.)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HeLa; HEK293.

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab83253 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB
ICC/IF
  • Application notesICC/IF: Use at a concentration of 1 µg/ml.
    WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 272 kDa (predicted molecular weight: 272 kDa).


    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • 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. Represses neuronal and myogenic differentiation. May enhance HIF1A function by sequestering HIF1AN away from HIF1A.
    • 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 aortic valve disease 1 (AOVD1) [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.
    • Post-translational
      modifications
      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). Following endocytosis, 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. Monoubiquitination at Lys-1759 is required for activation by gamma-secretase cleavage, it promotes interaction with AAK1, which stabilizes it. Deubiquitination by EIF3F is necessary for nuclear import of activated Notch.
      Hydroxylated at Asn-1955 by HIF1AN. Hydroxylated at Asn-2022 by HIF1AN (By similarity). Hydroxylation reduces affinity for HI1AN and may thus indirectly modulate negative regulation of NICD.
    • Cellular localizationCell membrane and Nucleus. Following proteolytical processing NICD is translocated to the nucleus.
    • Information by UniProt
    • Database links
    • Alternative names
      • hN1 antibody
      • Neurogenic locus Notch homolog protein 1 antibody
      • NICD antibody
      • NOTC1_HUMAN antibody
      • Notch 1 antibody
      • Notch 1 intracellular domain antibody
      • Notch homolog 1 translocation associated (Drosophila) antibody
      • notch1 antibody
      • TAN1 antibody
      • Translocation-associated notch protein TAN-1 antibody
      see all

    Anti-activated Notch1 antibody images

    • All lanes : Anti-activated Notch1 antibody (ab83253) at 1 µg/ml

      Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
      Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      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 : 272 kDa
      Additional bands at : 117 kDa,50 kDa. We are unsure as to the identity of these extra bands.

      Exposure time : 5 minutes
    • ICC/IF image of ab83253 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab83253, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa, HepG2 and MCF7 cells at 5µg/ml, and in 100% methanol fixed (5 min) HeLa, Hek293, HepG2 and MCF7 cells at 5µg/ml.

    References for Anti-activated Notch1 antibody (ab83253)

    ab83253 has not yet been referenced specifically in any publications.

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