Mouse Notch-1 Antibody

(2 citations)   
  • Species Reactivity
    Mouse
  • Specificity
    Detects mouse Notch-1 in direct ELISAs and Western blots. In direct ELISA, approximately 10% cross-reactivity with recombinant human Notch-1 and recombinant rat Notch-1 is observed and less than 1% cross-reactivity with recombinant mouse (rm) Notch-2 and rmNotch-3 is observed.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Chinese hamster ovary cell line CHO-derived recombinant mouse Notch‑1
    Ala19-Gln526
    Accession # AAM28905
  • 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
  • Immunohistochemistry
    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
Detection of Mouse Notch‑1 by Western Blot. Western blot shows lysates of bEnd.3 mouse endothelioma cell line. PVDF membrane was probed with 1 µg/mL of Mouse Notch‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5267) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Notch‑1 at approximately 300 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Immunohistochemistry
Notch‑1 in Mouse Neural Tube. Notch‑1 was detected in immersion fixed frozen sections of mouse neural tube (E13.5) using Mouse Notch‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5267) at 10 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red, upper panel; Catalog # NL010) and counterstained with DAPI (blue, lower panel). View our protocol for Fluorescent IHC Staining of Frozen 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

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). Mouse Notch-1 ECD aa 19‑526, which includes the first 13 EGF repeats, shows 94%, 91%, 86% and 79% aa identity with corresponding regions of rat, human, canine, and chicken Notch-1, respectively. This region also exhibits 55‑58% 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
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.

2 Citations: Showing 1 - 2
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Species
Applications
Sample Type
  1. Uncontrolled angiogenic precursor expansion causes coronary artery anomalies in mice lacking Pofut1
    Authors: Y Wang, B Wu, P Lu, D Zhang, B Wu, S Varshney, G Del Monte-, Z Zhuang, R Charafeddi, AH Kramer, NE Sibinga, NG Frangogian, RN Kitsis, RH Adams, K Alitalo, DJ Sharp, RP Harvey, P Stanley, B Zhou
    Nat Commun, 2017;8(1):578.
    Species: Hamster
    Sample Type: Whole Cells
    Application: Flow
  2. Antibodies that detect O-linked beta-D-N-acetylglucosamine on the extracellular domain of cell surface glycoproteins.
    Authors: Tashima Y, Stanley P
    J Biol Chem, 2014;289(16):11132-42.
    Species: Hamster
    Sample Type: Cell Lysates
    Application: WB
Cell and Tissue Staining Kits
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Anti-Sheep HRP-DAB Cell & Tissue Staining Kit

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Immunohistochemistry Reagents
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Antigen Retrieval Reagent-Acidic

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NorthernLights Guard Mounting Media

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VisUCyte Antigen Retrieval Reagent-Basic

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Isotype Controls
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Normal Sheep IgG Control

Ctrl 5-001-A 9
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Secondary Antibodies
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Sheep IgG HRP-conjugated Antibody

WB, Simple Western HAF016 14  
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Donkey Anti-Sheep IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL010 3
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Donkey Anti-Sheep IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL011 5
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Donkey Anti-Sheep IgG NorthernLights™ NL493-conjugated Antibody

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Donkey Anti-Sheep IgG Biotinylated Antibody

WB BAF016 2
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Sheep IgG (H+L) PE-conjugated Antibody

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Sheep IgG VisUCyte HRP Polymer Antibody

IHC VC006  
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Sheep IgG (H+L) APC-conjugated Antibody

Flow F0127  
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Donkey Anti-Sheep IgG (H+L) Fluorescein-conjugated Antibody

Flow F0125 1
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Donkey Anti-Sheep IgG (H+L) PerCP-conjugated Antibody

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