Rat Notch-2 Antibody

Catalog # Availability Size / Price Qty
AF1190
AF1190-SP
Notch‑2 in Rat Brain.
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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 as a 0.2 µm filtered solution in PBS.
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Label
Unconjugated

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.

Data Examples

Immunohistochemistry Notch‑2 in Rat Brain. View Larger

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 Notch‑2 in Embryonic Rat Liver. View Larger

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

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.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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

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 11th and 12thEGF-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
  1. Weinmaster, G. (2000) Curr. Opin. Genet. Dev. 10:363.
  2. Weinmaster, G. (1992) Development 116:931.
  3. Rebay, I. et al. (1991) Cell 67:687.
  4. Mumm, J.S. and R. Kopan (2000) Dev. Biol. 228:151.
  5. 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

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Cell and Tissue Staining Kits

Immunohistochemistry Reagents

Isotype Controls

Reconstitution Buffers

Secondary Antibodies

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