Rat Notch-2 Antibody

Catalog #: AF1190 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF1190
AF1190-SP

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Rat Notch-2 Antibody Summary

Species Reactivity

Rat

Specificity

Detects rat Notch-2 in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 5% cross-reactivity with recombinant rat Notch‑1 is observed, and less than 1% cross-reactivity with recombinant mouse Notch‑3 is observed.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

Mouse myeloma cell line NS0-derived recombinant rat Notch-2
Leu26-Glu492
Accession # Q9QW30

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.

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Applications

Recommended Concentration

Sample

Western Blot

0.1 µg/mL

Recombinant Rat Notch‑2 Fc Chimera (Catalog # 1190-NT)

Flow Cytometry

2.5 µg/10⁶ cells

Rat splenocytes

Immunohistochemistry

5-15 µg/mL

See below

Blockade of Receptor-ligand Interaction

In a functional ELISA, 2-5 µg/mL of this antibody will block 50% of the binding of 200 ng/mL of Recombinant Rat Jagged-1 Fc Chimera (Catalog # 599-JG) to immobilized Recombinant Rat Notch-2 Fc Chimera (Catalog # 1190‑NT) coated at 5 µg/mL (100 µL/well). At 20 μg/mL, this antibody will block >90% of the binding.

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

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

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Notch‑2 in Rat Brain. Notch‑2 was detected in perfusion fixed frozen sections of rat brain (choroid plexus) using 5 µg/mL Goat Anti-Rat Notch‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1190) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Immunohistochemistry

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Notch‑2 in Embryonic Rat Liver. Notch‑2 was detected in immersion fixed frozen sections of embryonic rat liver (15 d.p.c.) using 1.7 µg/mL Goat Anti-Rat Notch‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1190) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

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

Rat Notch-2 is a 300 kDa, type I transmembrane glycoprotein involved in a number of early-event developmental processes (1). In both vertebrates and invertebrates, Notch signaling is important for specifying cell fates and for defining boundaries between different cell types. The molecule is synthesized as a 2472 amino acid (aa) precursor that contains a putative 27 aa signal sequence, a 1650 aa extracellular region, a 23 aa transmembrane (TM) segment and a 772 aa cytoplasmic domain (2). The large Notch extracellular domain has 36 EGF-like repeats followed by three notch/Lin-12 repeats (LNR). Of the 36 EGF-like repeats, the 11^th and 12^thEGF-like repeats have been shown to be both necessary and sufficient for binding the ligands Serrate and Delta, in Drosophila (3). Cell surface Notch receptor is thought to be a heterodimer consisting of the ligand binding extracellular region associated with the remaining transmembrane protein, as a result of post-translational proteolytic cleavage by a furin-like enzyme. Upon ligand binding, additional proteolysis 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. In addition, an alternative Notch signaling pathway that is mediated by the full-length, uncleaved form of Notch-1 at the cell surface has been reported to suppress differentiation of myoblasts in response to ligand binding (5). Rat Notch-2 shows 92% and 95% aa identity to human and mouse Notch-2 extracellular domains, respectively. Relative to the extracellular region of rat Notch-1, rat Notch-2 exhibits 56% aa identity.

References

Weinmaster, G. (2000) Curr. Opin. Genet. Dev. 10:363.
Weinmaster, G. (1992) Development 116:931.
Rebay, I. et al. (1991) Cell 67:687.
Mumm, J.S. and R. Kopan (2000) Dev. Biol. 228:151.
Bush, G. et al. (2001) Dev. Biol. 229:494.

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