Mouse/Rat Notch-1 Antibody

(3 citations)   
  • Species Reactivity
    Mouse, Rat
  • Specificity
    Detects rat Notch‑1 in direct ELISAs and Western blots. In direct ELISAs and Western blots, less than 5% cross-reactivity with recombinant rat Notch-2 and recombinant mouse Notch-3 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant rat Notch‑1
    Arg20-Glu488 (Ala208Thr, Asp334Glu)
    Accession # Q07008
  • 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.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    Recombinant Rat Notch‑1 Fc Chimera (Catalog # 1057-TK)
  • Flow Cytometry
    2.5 µg/106 cells
    Rat cortical stem cells
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Blockade of Receptor-ligand Interaction
    In a functional ELISA, 1-3 μg/mL of Goat Anti-Mouse/Rat Notch-1Antibody (Catalog # AF1057) will block 50% of the binding of 200 ng/mL of RecombinantRat Jagged 1 Fc Chimera (Catalog # 599-JG)to immobilized Recombinant Rat Notch-1 Fc Chimera (Catalog # Catalog # 1057-TK) coated at5 μg/mL (100 μL/well). At 20 μg/mL, this antibody will block >80% of thebinding.
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    5-15 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
Immunocytochemistry
Notch‑1 in Rat Cortical Stem Cells. Notch‑1 was detected in immersion fixed undifferentiated rat cortical stem cells using Goat Anti-Mouse/Rat Notch‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1057) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 493-conjugated Anti-Goat IgG Secondary Antibody (green; Catalog # NL003) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.
Immunocytochemistry
Notch‑1 in Mouse Cortical Stem Cells. Notch‑1 was detected in immersion fixed undifferentiated mouse cortical stem cells using Goat Anti-Mouse/Rat Notch‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1057) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 493-conjugated Anti-Goat IgG Secondary Antibody (green; Catalog # NL003) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.
Immunohistochemistry
Notch‑1 in Rat Embryo. Notch‑1 was detected in immersion fixed paraffin-embedded sections of rat embryo (13 d.p.c.) using Goat Anti-Mouse/Rat Notch‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1057) at 10 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to labeling is in epidermis. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
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

Rat Notch-1 is a 300 kDa, type I transmembrane glycoprotein involved in a number of early-event developmental processes (1). In both vertebrates and invertebrates, Notch signaling is important for specifying cell fates and for defining boundaries between different cell types. The molecule is synthesized as a 2531 amino acid (aa) precursor that contains an 18 aa signal sequence, a 1705 aa extracellular region, a 23 aa transmembrane (TM) segment and a 785 aa cytoplasmic domain (2). The large Notch-1 extracellular domain has 36 EGF‑like repeats followed by three notch/Lin-12 repeats. Of the 36 EGF‑like repeats, the 11th and 12th EGF‑like repeats have been shown to be both necessary and sufficient for binding the ligands Delta and Serrate, in Drosophila (3). The Notch-1 cytoplasmic domain contains six ankyrin repeats, a glutamine-rich domain and a PEST sequence. The Notch-1 receptor undergoes post-translational proteolytic cleavage by a furin-like enzyme to form a heterodimer of the 1635 aa ligand binding extracellular region and the 877 aa transmembrane protein (4). Upon ligand binding, additional sequential proteolysis by TNF-converting enzyme and the Presenilin-dependent gamma -secretase results in the release of the Notch intracellular domain (NCID) which translocates into the nucleus where it functions as a transcription activator to initiate transcription of Notch-responsive genes (5). An alternative Notch signaling pathway that is mediated by the full-length form of Notch that has not been cleaved by the furin-like enzyme has also been reported (6). The rat Notch-1 extracellular domain shows 86% and 97% aa identity to human and mouse Notch-1 extracellular domains respectively. It also exhibits 56% and 50% aa identity with rat Notch-2 and Notch-3 extracellular domains, respectively.

  • References:
    1. Weinmaster, G. (2000) Curr. Opin. Genet. Dev. 10:363.
    2. Weinmaster, G. et al. (1991) Development 113:199.
    3. Rebay, I. et al. (1991) Cell 67:687.
    4. Rogeat, F. et al. (1998) Proc. Natl. Acad. Sci. USA 95:8108.
    5. Mumm, J.S. and R. Kopan (2000) Dev. Biol. 228:151.
    6. Bush, G. et al. (2001) Dev. Biol. 229:494.
  • 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
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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Species
Applications
Sample Type
  1. Macrophage-dependent tumor cell transendothelial migration is mediated by Notch1/Mena(INV)-initiated invadopodium formation
    Sci Rep, 2016;6(0):37874.
    Species: Human
    Sample Type: Whole Cells
    Application: Bioassay
  2. Human stem/progenitor cells from bone marrow enhance glial differentiation of rat neural stem cells: a role for transforming growth factor beta and Notch signaling.
    Authors: Robinson AP, Foraker JE, Ylostalo J, Prockop DJ
    Stem Cells Dev., 2011;20(2):289-300.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  3. Distinct stages of myelination regulated by gamma-secretase and astrocytes in a rapidly myelinating CNS coculture system.
    Authors: Watkins TA, Emery B, Mulinyawe S, Barres BA
    Neuron, 2008;60(4):555-69.
    Species: Mouse
    Sample Type: Whole Cells
    Application: ICC
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