Notch-3 Antibody [Unconjugated]

CD4- CD8- mouse thymocytes were stained with Rat Anti-Mouse CD25/IL-2R alpha PE‑conjugated Monoclonal Antibody (Catalog # FAB2438P) and either (A) Goat Anti-Mouse Notch‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1308) or (B) isotype control antibody (Catalog # AB-108-C) followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). View our protocol for Staining Membrane-associated Proteins.

Notch‑3 was detected in immersion fixed frozen sections of mouse embryo (E13) using Goat Anti-Mouse Notch‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1308) at 1 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the cell surface. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Notch‑3 was detected in immersion fixed frozen sections of mouse embryo (E13) using Goat Anti-Mouse Notch‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1308) at 3 µg/ml overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to the cell surface. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

• Reactivity: Mu
• Applications: WB, Flow, IHC, B/N, CyTOF-ready, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Notch-3 Antibody [Unconjugated] Summary

• Immunogen: S. frugiperda insect ovarian cell line Sf 21-derived recombinant mouse Notch-3
  Ala40-Glu468
  Accession # Q61982
• Specificity: Detects mouse Notch‑3 in direct ELISAs and Western blots. In direct ELISAs, less than 10% cross-reactivity with recombinant human
  Notch‑3 is observed and less than 5% cross-reactivity with recombinant rat (rr) Notch-1 and rrNotch-2 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: NOTCH3
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Blockade of Receptor-ligand Interaction
  • CyTOF-ready
  • Flow Cytometry 0.25 ug/10^6 cells
  • Immunofluorescence 1-15 ug/mL
  • Immunohistochemistry 1-15 ug/mL
  • Western Blot 0.1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
  *Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Notch-3 Antibody [Unconjugated]

• CADASIL
• CASILneurogenic locus notch homolog protein 3
• Notch (Drosophila) homolog 3
• notch 3Notch homolog 3 (Drosophila)
• Notch homolog 3
• Notch3
• Notch-3

Background

Mouse Notch-3 is part of the Notch family of type I transmembrane glycoproteins involved in a number of early-event developmental processes (1). The extracellular domain of Notch receptors interact with the extracellular domain of transmembrane ligands Jagged, Delta, and Serrate expressed on the surface of a neighboring cell. In both vertebrates and invertebrates, Notch signaling is important for specifying cell fates and for defining boundaries between different cell types. The Notch molecule is synthesized as a 2318 amino acid (aa) precursor that contains an 39 aa signal sequence, a 1603 aa extracellular region, a 20 aa transmembrane (TM) segment and a 655 aa cytoplasmic domain. The large Notch extracellular domain has 34 EGF-like repeats followed by three notch/Lin-12 repeats (LNR) (2). The 11^th and 12^th EGF-like repeats of Notch have been shown to be both necessary and sufficient for binding the ligands Serrate and Delta, in Drosophila (3). Notch-3 has the same biochemical mechanism of signal tranduction as Notch-1, where a series of cleavage events result in the release of the Notch intracellular domain (NICD). NICD translocates into the nucleus and initiates transcription of Notch-responsive genes (4). Thus Notch acts as both a ligand-binding receptor and a nuclear factor that regulates transcription.

Notch-3 is predominantly expressed in the developing central nervous system of mice (2). Mutations in Notch-3 in humans cause an autosomal dominant condition called CADASIL (cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy). This disorder is characterized by recurrent ischemic strokes at an early age without any underlying vascular risk and progressive dementia. Nearly all mutations leading to this disorder are clustered in the first 5 EGF repeats of the Notch-3 gene (5). Mouse Notch-3 shows 90% aa identity to human Notch-3 and 96% to rat Notch-3 over the entire protein.

1. Weinmaster, G. (2000) Curr. Opin. Genet. Dev. 10:363.
2. Lardelli, M. et al. (1994) Mech Dev. 46:123.
3. Rebay, I. et al. (1991) Cell 67:687.
4. Mizutani, T. et al. (2001) Proc. Natl. Acad. Sci. USA 98:9026.
5. Joutel, A. and E. Tounier-Lasserve (1998) Stem Cell & Dev. Biol. 9:619.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Notch-3 Antibody (AF1308)(9)

Publications using AF1308

• Meah, A;Vedarethinam, V;Bronstein, R;Gujarati, N;Jain, T;Mallipattu, SK;Li, Y;Wang, J; Single-Cell Spatial MIST for Versatile, Scalable Detection of Protein Markers Biosensors 2023-08-27 [PMID: 37754086] (IHC, Mouse)
• K Wei, I Korsunsky, JL Marshall, A Gao, GFM Watts, T Major, AP Croft, J Watts, PE Blazar, JK Lange, TS Thornhill, A Filer, K Raza, LT Donlin, CW Siebel, CD Buckley, S Raychaudhu, MB Brenner Notch signalling drives synovial fibroblast identity and arthritis pathology Nature, 2020-04-22;582(7811):259-264. 2020-04-22 [PMID: 32499639] (IHC, Mouse)
• R Diéguez-Hu, K Kato, BD Giaimo, M Nieminen-K, H Arf, F Ferrante, M Bartkuhn, T Zimmermann, MG Bixel, HM Eilken, S Adams, T Borggrefe, P Vajkoczy, RH Adams Loss of the transcription factor RBPJ induces disease-promoting properties in brain pericytes Nat Commun, 2019-06-27;10(1):2817. 2019-06-27 [PMID: 31249304] (IHC, Mouse)
• F Ferrandino, G Bernardini, G Tsaouli, P Grazioli, AF Campese, C Noce, A Ciuffetta, A Vacca, ZM Besharat, D Bellavia, I Screpanti, MP Felli Intrathymic Notch3 and CXCR4 combinatorial interplay facilitates T-cell leukemia propagation Oncogene, 2018-07-23;0(0):. 7/23/2018 [PMID: 30038265]
• K Kato, R Diéguez-Hu, DY Park, SP Hong, S Kato-Azuma, S Adams, M Stehling, B Trappmann, JL Wrana, GY Koh, RH Adams Pulmonary pericytes regulate lung morphogenesis Nat Commun, 2018-06-22;9(1):2448. 2018-06-22 [PMID: 29934496] (IHC-P, Mouse)
• Y Zhang, S Dong, J Wang, M Wang, M Chen, H Huang Involvement of Notch2 in all?trans retinoic acid?induced inhibition of mouse embryonic palate mesenchymal cell proliferation Mol Med Rep, 2017-07-06;16(3):2538-2546. 2017-07-06 [PMID: 28713967] (IHC, Western Blot, Mouse)
• Prolyl-isomerase Pin1 controls Notch3 protein expression and regulates T-ALL progression Oncogene, 2016-02-15;35(36):4741-51. 2/15/2016 [PMID: 26876201]
• Anupama Gopisetty, Palash Bhattacharya, Christine Haddad, Joseph C. Bruno, Chenthamarakshan Vasu, Lucio Miele et al. OX40L/Jagged1 Cosignaling by GM-CSF–Induced Bone Marrow-Derived Dendritic Cells Is Required for the Expansion of Functional Regulatory T Cells The Journal of Immunology 2013-06-01 [PMID: 23630352]
• Blanpain C, Lowry WE, Pasolli HA, Fuchs E Canonical notch signaling functions as a commitment switch in the epidermal lineage. Genes Dev., 2006-11-01;20(21):3022-35. 2006-11-01 [PMID: 17079689] (IHC-P, Mouse)

Bioinformatics

• Gene Symbol: NOTCH3
• Uniprot:
  • Q61982()