Mouse Notch-1 Antibody MAB5267: R&D Systems

Mouse Notch-1 Antibody

  
  • Species Reactivity
    Mouse
  • Specificity
    Detects mouse Notch-1 intracellular domain in direct ELISAs.
  • Source
    Monoclonal Mouse IgG1 Clone # N1A
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Mouse embryonic fibroblast cell line PA317-derived recombinant mouse Notch-1
    Accession # Q01705
  • 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
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Intracellular Staining by Flow Cytometry
    0.25 µg/106 cells
    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
Intracellular Staining by Flow Cytometry
Detection of Notch‑1 in RPMI 8226 Human Cell Line by Flow Cytometry. RPMI 8226 human multiple myeloma cell line was stained with Mouse Notch‑1 Monoclonal Antibody (Catalog # MAB5267, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')2 Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 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

Notch-1 is a 300 kDa type I transmembrane glycoprotein that is one of four 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, it promotes T cell over B cell development from a common precursor (2). Mouse Notch-1 is synthesized as a 2531 amino acid (aa) precursor that contains an 18 aa signal sequence, a 1707 aa extracellular domain (ECD) with 36 EGF-like repeats and three Lin-12/notch repeats (LNR), a 21 aa transmembrane (TM) segment and a 785 aa cytoplasmic domain that contains six ankyrin repeats, a glutamine-rich domain and a PEST sequence. The 11th and 12th EGF-like repeats, that bind ligands such as Jagged and Delta-like families in humans, correspond to aa 412-488 in mouse Notch-1 (6). Elongation of O-linked fucose chains 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). The Notch-1 receptor undergoes post-translational furin-type proteolytic cleavage, generating a heterodimer through the interaction of a hydrophobic area C-terminal to the LNR on the extracellular region with the transmembrane/cytoplasmic portion (8, 9). Upon ligand binding, additional sequential proteolysis by TNF-converting enzyme (ADAM17) 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).

  • 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
Related Research Areas
Isotype Controls
Description Application Cat# Citations Images  

Mouse IgG1 Isotype Control

Ctrl MAB002 138  
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Mouse IgG1 Isotype Control

Ctrl MAB002R 1
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Secondary Antibodies
Description Application Cat# Citations Images  

Mouse IgG HRP-conjugated Antibody

WB HAF007 26  
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Mouse F(ab)2 IgG (H+L) PE-conjugated Antibody

Flow F0102B 10  
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Donkey Anti-Mouse IgG NorthernLights™ NL557-conjugate Antibody

Flow, IHC, ICC NL007 9
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Donkey Anti-Mouse IgG Biotinylated Antibody

WB BAF018 1
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Mouse IgG HRP-conjugated Antibody

WB HAF018 9  
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Mouse F(ab)2 IgG (H+L) APC-conjugated Antibody

Flow F0101B 2  
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Donkey Anti-Mouse IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL008 8
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Donkey Anti-Mouse IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL009 5
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Goat Anti-Mouse IgG Biotinylated Antibody

WB BAF007 4
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Mouse IgG Antibody

WB AF007 4
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Mouse F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

Flow F0103B 5  
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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
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Mouse IgG VisUCyte HRP Polymer Antibody

IHC VC001  
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Goat Anti-Mouse IgG Fc Affinity Purified Polyclonal Ab

G-202-C 2
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Mouse F(ab)2 IgG (H+L) PerCP-conjugated Antibody

Flow F0114  
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