In a functional ELISA, 1.5-6 µg/mL of this antibody will block 50% of the binding of 200 ng/mL of Recombinant Human beta -NGF (Catalog # 256-GF) to immobilized Recombinant Mouse Sortilin (Catalog # 2934-ST) coated at 4 µg/mL (100 µL/well). At 100 μg/mL, this antibody will block >90% of the binding.
Please Note: Optimal dilutions should be determined by each laboratory for each application.
are available in the Technical Information section on our website.
Detection of Mouse Sortilin by Western Blot.
Western blot shows lysates of mouse brain (total) tissue, mouse brain (cerebellum) tissue, and mouse brain (cortex) tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse Sortilin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2934) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Sortilin at approximately 110 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Sortilin in Mouse Brain.
Sortilin was detected in perfusion fixed frozen sections of mouse brain (rostral ventral medulla) using Goat Anti-Mouse Sortilin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2934) at 1.7 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). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.
Detection of Mouse Sortilin by Simple WesternTM.
Simple Western lane view shows lysates of mouse brain (cortex) tissue, loaded at 0.2 mg/mL. A specific band was detected for Sortilin at approximately 122 kDa (as indicated) using 10 µg/mL of Goat Anti-Mouse Sortilin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2934) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
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.
Sortilin (neurotensin receptor 3, glycoprotein 95) is a 95 kDa Type I transmembrane monomeric glycoprotein that is one of five known members of the mammalian vacuolar protein sorting 10p domain (Vps10p-D) family of sorting receptors (1, 2). Mouse preprosortilin is processed by signal sequence cleavage followed by propeptide cleavage at a furin recognition site. The cationic propeptide exhibits pH-dependent high affinity binding that blocks the Sortilin ligand binding site both pre‑ and post-cleavage (3). The extracellular/luminal sequence comprises the Vps10p domain, including 10 conserved cysteines (10CC) essential for ligand binding (2). The cytoplasmic domain sorting motifs confer all trafficking during synthesis, targeting to lysosomes, endocytosis and Golgi-endosome transport; as little as 10% may be found on the cell surface (4). Mature mouse Sortilin shares 98% amino acid (aa) identity with rat, and 91% aa identity with human and canine sortilin. During murine development, sortilin is mainly expressed in the nervous system (5), where it is a receptor for neuropeptides including neurotensin, nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) (6‑9). ProNGF (or the NGF propeptide alone) binds sortilin with a much higher affinity (KD ~5-8 nM) than does mature NGF (KD ~90 nM). The complex of sortilin, pro‑NGF and the receptor p75ntr results in endocytosis of proNGF and induction of apoptosis (7). Similar results have been obtained with proBDNF and BDNF (8‑9). Sortilin is expressed in other tissues including testis, skeletal muscle and fat (1, 10). It is essential and sufficient for biogenesis of Glut4 storage vesicles necessary for insulin responsiveness in adipocytes (10). Sortilin also binds lipoprotein lipase (11), apoE (2) and RAP (1, 11). Binding is competitive, indicating that although unrelated, targets likely bind the same site.
Petersen, C.M. et al. (1997) J. Biol. Chem. 272:3599.
Westergaard, U.B. et al. (2004) J. Biol. Chem. 279:50221.
Petersen, C.M. et al. (1998) EMBO J. 18:595.
Nielsen, M.S. et al. (2001) EMBO J. 20:2180.
Hermans-Borgmeyer, I. et al. (1999) Mol. Brain Res. 65:216.
Mazella, J. et al. (1998) J. Biol. Chem. 273:26273.
Nykjaer, A et al. (2004) Nature 427:843.
Teng, H.K. et al. (2005) J. Neurosci. 25:5455.
Chen, Z.-Y. et al. (2004) J. Neurosci. 25:6156.
Shi, J and K.V. Kandror (2005) Dev. Cell 9:99.
Nielsen, M.S. et al. (1999) J. Biol. Chem. 274:8832.
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.
Submit a review and receive a $25US/€18/£15/$25CAN amazon gift card if you include an image - $10US/€7/£6/$10CAN Amazon card for reviews without an image. Limited to verified customers in USA, Canada and Europe.