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Mouse Notch-1 Antibody

Catalog #: AF5267
13 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
AF5267
AF5267-SP
Detection of Mouse Notch‑1 by Western Blot.
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Citations (13)
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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Mouse Notch-1 Antibody Summary

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 either lyophilized or 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.

Scientific Data

Western Blot Detection of Mouse Notch‑1 antibody by Western Blot. View Larger

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 antibody in Mouse Neural Tube by Immunohistochemistry (IHC-Fr). View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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

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Citations for Mouse Notch-1 Antibody

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.

13 Citations: Showing 1 - 10
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  1. O-GlcNAc on NOTCH1 EGF repeats regulates ligand-induced Notch signaling and vascular development in mammals
    Authors: Sawaguchi S, Varshney S, Ogawa M et al.
    eLife
  2. Cell-Cell Contact Area Affects Notch Signaling and Notch-Dependent Patterning
    Authors: O Shaya, U Binshtok, M Hersch, D Rivkin, S Weinreb, L Amir-Zilbe, B Khamaisi, O Oppenheim, RA Desai, RJ Goodyear, GP Richardson, CS Chen, D Sprinzak
    Dev. Cell, 2017-03-13;40(5):505-511.e6.
  3. In vivo evidence for GDP-fucose transport in the absence of transporter SLC35C1 and putative transporter SLC35C2
    Authors: Lu, L;Varshney, S;Yuan, Y;Wei, HX;Tanwar, A;Sundaram, S;Nauman, M;Haltiwanger, RS;Stanley, P;
    The Journal of biological chemistry
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  4. EOGT enables residual Notch signaling in mouse intestinal cells lacking POFUT1
    Authors: Nauman, M;Varshney, S;Choi, J;Augenlicht, LH;Stanley, P;
    Scientific reports
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  5. Fringe GlcNAc-transferases differentially extend O-fucose on endogenous NOTCH1 in mouse activated T cells
    Authors: K Matsumoto, V Kumar, S Varshney, AV Nairn, A Ito, F Pennarubia, KW Moremen, P Stanley, RS Haltiwange
    The Journal of Biological Chemistry, 2022-05-25;0(0):102064.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  6. Canonical Notch ligands and Fringes have distinct effects on NOTCH1 and NOTCH2
    Authors: S Kakuda, RK Lopilato, A Ito, RS Haltiwange
    J. Biol. Chem., 2020-08-19;0(0):.
    Species: Human
    Sample Types: Cell Lysates, Whole Cells
    Applications: Flow Cytometry, Western Blot
  7. Notch signalling defines dorsal root ganglia neuroglial fate choice during early neural crest cell migration
    Authors: S Wiszniak, Q Schwarz
    BMC Neurosci, 2019-04-29;20(1):21.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC-F
  8. Two novel protein O-glucosyltransferases that modify sites distinct from POGLUT1 and affect Notch trafficking and signaling
    Authors: H Takeuchi, M Schneider, DB Williamson, A Ito, M Takeuchi, PA Handford, RS Haltiwange
    Proc. Natl. Acad. Sci. U.S.A., 2018-08-20;0(0):.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  9. 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, 2017-09-18;8(1):578.
    Species: Hamster
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  10. O-GlcNAc on NOTCH1 EGF repeats regulates ligand-induced Notch signaling and vascular development in mammals
    Authors: Sawaguchi S, Varshney S, Ogawa M et al.
    eLife
  11. 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, 2014-02-26;289(16):11132-42.
    Species: Hamster
    Sample Types: Cell Lysates
    Applications: Western Blot
  12. Inhibition of Delta-induced Notch signaling using fucose analogs
    Authors: M Schneider, V Kumar, LU Nordstrøm, L Feng, H Takeuchi, H Hao, VC Luca, KC Garcia, P Stanley, P Wu, RS Haltiwange
    Nat. Chem. Biol., 2017-11-27;0(0):.
  13. Lunatic, Manic, and Radical Fringe Each Promote T and B Cell Development
    Authors: Yinghui Song, Vivek Kumar, Hua-Xing Wei, Ju Qiu, Pamela Stanley
    The Journal of Immunology

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