Recombinant Rat Notch‑1 Fc Chimera (Catalog # 1057-TK)
2.5 µg/106 cells
Rat cortical stem cells
Blockade of Receptor-ligand Interaction
In a functional ELISA, 1-3 μg/mL of Goat Anti-Mouse/Rat Notch-1Antibody (Catalog # AF1057) will block 50% of the binding of 200 ng/mL of RecombinantRat Jagged 1 Fc Chimera (Catalog # 599-JG)to immobilized Recombinant Rat Notch-1 Fc Chimera (Catalog # Catalog # 1057-TK) coated at5 μg/mL (100 μL/well). At 20 μg/mL, this antibody will block >80% of thebinding.
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.
are available in the Technical Information section on our website.
Notch‑1 in Rat Cortical Stem Cells.
Notch‑1 was detected in immersion fixed undifferentiated rat cortical stem cells using Goat Anti-Mouse/Rat Notch‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1057) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 493-conjugated Anti-Goat IgG Secondary Antibody (green; Catalog # NL003) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.
Notch‑1 in Mouse Cortical Stem Cells.
Notch‑1 was detected in immersion fixed undifferentiated mouse cortical stem cells using Goat Anti-Mouse/Rat Notch‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1057) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 493-conjugated Anti-Goat IgG Secondary Antibody (green; Catalog # NL003) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.
Notch‑1 in Rat Embryo.
Notch‑1 was detected in immersion fixed paraffin-embedded sections of rat embryo (13 d.p.c.) using Goat Anti-Mouse/Rat Notch‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1057) at 10 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to labeling is in epidermis. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
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.
Rat Notch-1 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 2531 amino acid (aa) precursor that contains an 18 aa signal sequence, a 1705 aa extracellular region, a 23 aa transmembrane (TM) segment and a 785 aa cytoplasmic domain (2). The large Notch-1 extracellular domain has 36 EGF‑like repeats followed by three notch/Lin-12 repeats. Of the 36 EGF‑like repeats, the 11th and 12th EGF‑like repeats have been shown to be both necessary and sufficient for binding the ligands Delta and Serrate, in Drosophila (3). The Notch-1 cytoplasmic domain contains six ankyrin repeats, a glutamine-rich domain and a PEST sequence. The Notch-1 receptor undergoes post-translational proteolytic cleavage by a furin-like enzyme to form a heterodimer of the 1635 aa ligand binding extracellular region and the 877 aa transmembrane protein (4). Upon ligand binding, additional sequential proteolysis by TNF-converting enzyme and the Presenilin-dependent gamma -secretase results in the release of the Notch intracellular domain (NCID) which translocates into the nucleus where it functions as a transcription activator to initiate transcription of Notch-responsive genes (5). An alternative Notch signaling pathway that is mediated by the full-length form of Notch that has not been cleaved by the furin-like enzyme has also been reported (6). The rat Notch-1 extracellular domain shows 86% and 97% aa identity to human and mouse Notch-1 extracellular domains respectively. It also exhibits 56% and 50% aa identity with rat Notch-2 and Notch-3 extracellular domains, respectively.
Weinmaster, G. (2000) Curr. Opin. Genet. Dev. 10:363.
Weinmaster, G. et al. (1991) Development 113:199.
Rebay, I. et al. (1991) Cell 67:687.
Rogeat, F. et al. (1998) Proc. Natl. Acad. Sci. USA 95:8108.
Mumm, J.S. and R. Kopan (2000) Dev. Biol. 228:151.
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