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Human Notch-1 Antibody

Catalog #: MAB5317
1 Citation Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
MAB5317-100
MAB5317-500
MAB5317-SP
Detection of Notch-1 in U2OS Human Cell Line by Flow Cytometry.
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Product Details
Citations (1)
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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human Notch-1 Antibody Summary

Species Reactivity
Human
Specificity
Detects human Notch-1.
Source
Monoclonal Mouse IgG1 Clone # 527425
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Mouse myeloma cell line NS0-derived recombinant human Notch-1
aa 19-526
Accession # P46531
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
Flow Cytometry
0.25 µg/106 cells
U2OS human osteosarcoma cell line and Molt-4 cells
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 

Human Notch-1 Sandwich Immunoassay

Recommended Concentration
Reagent
ELISA Capture (Matched Antibody Pair)
2-8 µg/mL 

Use in combination with:

Detection Reagent: Human Notch‑1 Biotinylated Antibody (Catalog # BAM53171)

Standard: Recombinant Human Notch-1 Fc Chimera Protein, CF (Catalog # 3647-TK)

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

Flow Cytometry Detection of Notch-1 antibody in U2OS Human Cell Line antibody by Flow Cytometry. View Larger

Detection of Notch-1 in U2OS Human Cell Line by Flow Cytometry. U2OS human osteosarcoma cell line was stained with Human Notch‑1 Monoclonal Antibody (Catalog # MAB5317, filled histogram) or isotype control antibody (MAB002, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG F(ab')2Secondary Antibody (F0101B).

Flow Cytometry View Larger

Detection of Notch‑1 in Molt-4 cells by Flow Cytometry. Molt-4 cells were stained with Mouse Anti-Human Notch‑1 Monoclonal Antibody (Catalog # MAB5317, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). View our protocol for Staining Membrane-associated Proteins.

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 0.5 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

Human Notch-1 is a 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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Citation for Human 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.

1 Citation: Showing 1 - 1

  1. The CD46-Jagged1 interaction is critical for human TH1 immunity.
    Authors: Le Friec G, Sheppard D, Whiteman P, Karsten C, Shamoun S, Laing A, Bugeon L, Dallman M, Melchionna T, Chillakuri C, Smith R, Drouet C, Couzi L, Fremeaux-Bacchi V, Kohl J, Waddington S, McDonnell J, Baker A, Handford P, Lea S, Kemper C
    Nat Immunol, 2012-10-21;13(12):1213-21.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry

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