Human Notch-1 Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human Notch-1 in direct ELISAs and Western blots. In direct ELISAs, approximately 40% cross‑reactivity with with recombinant mouse Notch-1 (aa 19‑526) and recombinant rat Notch-1 (aa 20‑488) is observed and less than 1% cross-reactivity with recombinant human (rh) Notch-2 and rhNotch-3 is observed.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human Notch-1
    Ala19-Gln526
    Accession # P46531
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human Notch‑1 by Western Blot. Western blot shows lysates of Nalm‑6 human Pre-B acute lymphocytic leukemia cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Notch‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5317) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Notch‑1 at approximately 140 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Notch-1

Human Notch-1 is a 120-300 kDa type I transmembrane glycoprotein that is one of four human Notch homologues involved in developmental processes (1‑3). Notch signaling is important for maintaining stem cells and inducing differentiation, especially in the nervous system and lymphoid tissues (2‑4). Notch can specify binary cell fates; for example, promoting T- over B-cell development from a common precursor (2). More than 50% of human T-lineage acute lymphoblastic leukemia (T‑ALL) have activating mutations of Notch1 (1, 5). Human Notch-1 is synthesized as a 2556 amino acid (aa) precursor that contains an 18 aa signal sequence, a 1718 aa extracellular domain (ECD) with 36 EGF-like repeats and three Lin-12/Notch repeats (LNR), a 23 aa transmembrane (TM) segment and a 785 aa cytoplasmic domain containing six ankyrin repeats, a glutamine-rich domain and a PEST sequence. The 11th and 12th EGF-like repeats bind ligands including Jagged and Delta-like families in humans (6). O-fucosylation by Fringe family members at a site within this region can inhibit the interaction of Notch with Jagged ligands, thereby promoting Delta-like ligand interactions (7). Notch-1 receptor undergoes post-translational furin-type proteolytic cleavage, forming a heterodimer through interaction of a hydrophobic area C-terminal to the LNR on the 1647 aa ligand-binding extracellular region with the 891 aa transmembrane/cytoplasmic portion (8, 9). Upon ligand binding, additional sequential proteolysis by TNF-converting enzyme (ADAM-17) and the presenilin-dependent gamma -secretase results in the release of the Notch intracellular domain (NICD) which translocates into the nucleus, activating transcription of Notch-responsive genes (10). Human Notch-1 ECD aa 19‑526, including the first 13 EGF repeats, shows 91% aa identity with corresponding regions of mouse and rat, 89% with canine, and 79% with chicken Notch-1. This region also exhibits 60% aa identity with human Notch-2 and Notch-3.

  • References:
    1. Ellisen, L. W. et al. (1991) Cell 66:649.
    2. Dumortier, A. et al. (2005) Int. J. Hematol. 82:277.
    3. Yoon, K. and N. Gaiano (2005) Nat. Neurosci. 8:709.
    4. Androutsellis-Theotokis, A. et al. (2006) Nature 442:823.
    5. Weng, A. P. et al. (2004) Science 306:269.
    6. Hambleton, S. et al. (2004) Structure 12:2173.
    7. Yang, L. et al. (2005) Mol. Biol. Cell 16:927.
    8. Sanchez-Irizarry, C. et al. (2004) Mol. Cell. Biol. 24:9265.
    9. Logeat, F. et al. (1998) Proc. Natl. Acad. Sci. USA 95:8108.
    10. Mumm, J.S. and R. Kopan (2000) Dev. Biol. 228:151.
  • Entrez Gene IDs:
    4851 (Human); 25496 (Rat)
  • Alternate Names:
    EC 2.1.2.11; EC 3.4.21.68; hN1; Notch (Drosophila) homolog 1 (translocation-associated); notch 1Notch homolog 1, translocation-associated (Drosophila); Notch homolog 1, translocation-associated; Notch1; Notch-1; TAN1; TAN1neurogenic locus notch homolog protein 1; Translocation-associated notch protein TAN-1
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Sheep IgG (H+L) APC-conjugated Antibody

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Donkey Anti-Sheep IgG (H+L) Fluorescein-conjugated Antibody

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Donkey Anti-Sheep IgG (H+L) PerCP-conjugated Antibody

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