Mouse/Rat Notch-2 Antibody AF5196: R&D Systems

Mouse/Rat Notch-2 Antibody

  
  • Species Reactivity
    Mouse, Rat
  • Specificity
    Detects mouse and rat Notch‑2 in direct ELISAs and Western blots. In direct ELISAs, approximately 50% cross‑reactivity with recombinant human Notch-2 is observed and less than 1% cross-reactivity with recombinant mouse (rm) Notch-1 and rmNotch-3 is observed.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Chinese hamster ovary cell line CHO-derived recombinant mouse Notch‑2
    Leu26-Val528
    Accession # NP_035058
  • 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
    2 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 and Rat Notch‑2 by Western Blot. Western blot shows lysates of Balb/3T3 mouse embryonic fibroblast cell line and rat embryonic hippocampal neuron tissue. PVDF Membrane was probed with 2 µg/mL of Sheep Anti-Mouse/Rat Notch‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5196) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Notch‑2 at approximately 275 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1 .
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-2

Notch-2 is a 300 kDa type I transmembrane glycoprotein that is one of four Notch homologues involved in development (1‑3). Although Notch proteins are structurally and functionally similar, deletion of either Notch-1 or Notch-2 is lethal, demonstrating that not all functions overlap (4, 5). Mice with hypomorphic Notch-2 show defects in development of kidney, heart and eye (6). Notch-2 is upregulated in mature B cells and is critical for differentiation to splenic marginal zone B cells (7). Notch-2 is also preferentially expressed in choroid plexus epithelia and neuronal precursors (8, 9). Mouse Notch-2 is synthesized as a 2470 amino acid (aa) precursor that contains a 25 aa signal sequence, a 1652 aa extracellular domain (ECD), a 21 aa transmembrane (TM) segment, and a 772 aa cytoplasmic domain. The ECD contains 35 EGF‑like repeats and three Lin-12/Notch repeats, while the cytoplasmic region shows six ankyrin repeats, a glutamine‑rich domain and a PEST sequence. Binding of ligands, including Jagged and Delta-like molecules, has been localized to the 11th and 12th EGF-like repeats of human Notch-1 (corresponding to aa 413‑492 in mouse Notch-2) (10). Notch receptors undergo post‑translational furin-type proteolytic cleavage (11). This forms a heterodimer through the interaction of a hydrophobic area in the ligand‑binding extracellular region with the TM/cytoplasmic portion (11, 12). 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 (13). Mouse Notch-2 ECD (aa 26‑528) shares 93%, 96%, 92% and 92% aa identity with the corresponding regions of human, rat, canine, and bovine Notch-2, respectively. This region also exhibits 55‑58% aa identity with mouse Notch-1 and Notch-3.

  • References:
    1. Lardelli, M. and U. Lendahl (1993) Exp. Cell Res. 204:364.
    2. Dumortier, A. et al. (2005) Int. J. Hematol. 82:277.
    3. Yoon, K. and N. Gaiano (2005) Nat. Neurosci. 8:709.
    4. Hamada, Y. et al. (1999) Development 126:3415.
    5. Shimizu, K. et al. (2002) Biochem. Biophys. Res. Commun. 291:775.
    6. McCright, B. et al. (2001) Development 128:491.
    7. Saito, T. et al. (2003) Immunity 18:675.
    8. Irvin, D. K. et al. (2001) J. Comp. Neurol. 436:167.
    9. Solecki, D. J. et al. (2001) Neuron 31:557.
    10. Hambleton, S. et al. (2004) Structure 12:2173.
    11. Logeat, F. et al. (1998) Proc. Natl. Acad. Sci. USA 95:8108.
    12. Sanchez-Irizarry, C. et al. (2004) Mol. Cell. Biol. 24:9265.
    13. Mumm, J.S. and R. Kopan (2000) Dev. Biol. 228:151.
  • Entrez Gene IDs:
    4853 (Human); 29492 (Rat)
  • Alternate Names:
    AGS2; hN2; neurogenic locus notch homolog protein 2; Notch (Drosophila) homolog 2; notch 2Notch homolog 2 (Drosophila); Notch homolog 2; Notch2; Notch-2
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