Home / Notch-2 / Human Notch-2 Intracellular Domain Antibody

Human Notch-2 Intracellular Domain Antibody

Catalog #: AF3735
6 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
AF3735
AF3735-SP
Detection of Notch‑2 in Human Kidney.
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Citations (6)
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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human Notch-2 Intracellular Domain Antibody Summary

Species Reactivity
Human
Specificity
Detects human Notch-2 Intracellular Domain in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 5% cross‑reactivity with recombinant human (rh) Notch-3 (aa 2195‑2321), rhNotch-6 (aa 2251‑2556) and rhNotch-4 (aa 1778‑2003) is observed.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant human Notch‑2
Asp2063-Pro2413
Accession # Q04721
Formulation
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.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.1 µg/mL
Recombinant Human Notch‑2 Intracellular Domain
Immunohistochemistry
5-15 µg/mL
Immersion fixed paraffin-embedded sections of human kidney

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

Immunohistochemistry View Larger

Detection of Notch‑2 in Human Kidney. Notch‑2 was detected in immersion fixed paraffin-embedded sections of human kidney using Goat Anti-Human Notch‑2 Intracellular Domain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3735) at 5 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004) or the HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the cytoplasm and nucleus. View our protocol for Chromogenic IHC Staining of Paraffin-embedded 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. For liquid material, refer to CoA for concentration.
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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-2

Human Notch-2 is a 300 kDa type I transmembrane glycoprotein that is one of four human Notch homologues involved in developmental processes (1‑3). Although Notch proteins are structurally and functionally similar, deletion of either Notch-1 or Notch-2 is lethal, showing that not all functions overlap (4, 5). Mice with hypomorphic Notch-2 show defects in the development of kidney, heart and eyes (6). Notch-2 is up‑regulated 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). Human Notch-2 is synthesized as a 2471 amino acid (aa) precursor that contains a 25 aa signal sequence, a 1652 aa extracellular domain (ECD), and a 794 aa transmembrane (TM) and cytoplasmic segment. 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 in humans, has been localized to the 11th and 12th EGF-like repeats of Notch (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). Human Notch-2 ECD (aa 26‑530) shows 93%, 93%, 96% and 96% aa identity with the corresponding regions of mouse, rat, canine, and bovine Notch-2, respectively. This region also exhibits approximately 60% aa identity with human Notch-1 and Notch-3.

References
  1. Stifani, S. et al. (1992) Nat. Genet. 2:119.
  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. Comm. 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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Citations for Human Notch-2 Intracellular Domain 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.

6 Citations: Showing 1 - 6
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  1. Cooperation Between Cancer and Fibroblasts in Vascular Mimicry and N2-Type Neutrophil Recruitment via Notch2–Jagged1 Interaction in Lung Cancer
    Authors: Ying-Ming Tsai, Kuan-Li Wu, Yu-Wei Liu, Wei-An Chang, Yung-Chi Huang, Chao-Yuan Chang et al.
    Frontiers in Oncology
  2. A role for Notch signaling in trophoblast endovascular invasion and in the pathogenesis of pre-eclampsia
    Authors: Nathan M. Hunkapiller, Malgorzata Gasperowicz, Mirhan Kapidzic, Vicki Plaks, Emin Maltepe, Jan Kitajewski et al.
    Development
  3. NOTCH2 negatively regulates metastasis and epithelial-Mesenchymal transition via TRAF6/AKT in nasopharyngeal carcinoma
    Authors: Y Zou, R Yang, ML Huang, YG Kong, JF Sheng, ZZ Tao, L Gao, SM Chen
    J. Exp. Clin. Cancer Res., 2019-11-07;38(1):456.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  4. Involvement of Notch in activation and effector functions of gammadelta T cells.
    Authors: Gogoi D, Dar A, Chiplunkar S
    J Immunol, 2014-01-31;192(5):2054-62.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  5. Influence of the Notch system in the therapeutic response of American tegumentary leishmaniasis.
    Authors: Rodrigues KM, Oliveira MP, Maretti-Mira AC, Oliveira-Neto MP, Mattos MS, Silva L, Soares DA, Dolci EL, Perico RA, Pirmez C
    Br. J. Dermatol., 2011-05-17;164(6):1228-34.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-Fr
  6. Delineation of a cellular hierarchy in lung cancer reveals an oncofetal antigen expressed on tumor-initiating cells.
    Authors: Damelin M, Geles KG, Follettie MT, Yuan P, Baxter M, Golas J, Dijoseph JF, Karnoub M, Huang S, Diesl V, Behrens C, Choe SE, Rios C, Gruzas J, Sridharan L, Dougher M, Kunz A, Hamann PR, Evans D, Armellino D, Khandke K, Marquette K, Tchistiakova L, Boghaert ER, Abraham RT, Wistuba II, Zhou BB
    Cancer Res., 2011-05-03;71(12):4236-46.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-Fr

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