Human Sortilin Antibody

(1 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human and mouse Sortilin in direct ELISAs and Western blots.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human Sortilin
    Ser78-Asn755
    Accession # Q99523
  • 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
    2 µg/mL
    See below
  • Simple Western
    10 µg/mL
    See below
  • Flow Cytometry
    0.25 µg/106 cells
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Blockade of Receptor-ligand Interaction
    In a functional ELISA, 3-9 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 Human Sortilin (Catalog # 3154-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. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human Sortilin by Western Blot. Western blot shows lysates of human motor cortex. PVDF membrane was probed with 2 µg/mL of Goat Anti-Human Sortilin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3154) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Sortilin at approximately 95-105 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Sortilin in Human Blood Monocytes by Flow Cytometry. Human peripheral blood monocytes were stained with Goat Anti-Human Sortilin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3154, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). View our protocol for Staining Membrane-associated Proteins.
Detection of Sortilin in K562 Human Cell Line by Flow Cytometry. K562 human chronic myelogenous leukemia cell line was stained with Goat Anti-Human Sortilin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3154, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). View our protocol for Staining Membrane-associated Proteins.
Immunohistochemistry
Sortilin in Human Brain. Sortilin was detected in immersion fixed paraffin-embedded sections of human brain using Goat Anti-Human Sortilin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3154) 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 Paraffin-embedded Tissue Sections.
Detection of Human Sortilin by Simple WesternTM. Simple Western lane view shows lysates of human brain (cerebellum) tissue, loaded at 0.2 mg/mL. A specific band was detected for Sortilin at approximately 82 kDa (as indicated) using 10 µg/mL of Goat Anti-Human Sortilin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3154). This experiment was conducted under reducing conditions and using the
12-230 kDa separation system.        
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • 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: Sortilin

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). Human 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 (10 CC) 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 human Sortilin shares 91% aa identity with mouse and rat Sortilin and 93% aa identity with dog. 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 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 pro-BDNF 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.

  • References:
    1. Petersen, C.M. et al. (1997) J. Biol. Chem. 272:3599.
    2. Westergaard, U.B. et al. (2004) J. Biol. Chem. 279:50221.
    3. Petersen, C.M. et al. (1998) EMBO J. 18:595.
    4. Nielsen, M.S. et al. (2001) EMBO J. 20:2180.
    5. Hermans-Borgmeyer, I. et al. (1999) Mol. Brain Res. 65:216.
    6. Mazella, J. et al. (1998) J. Biol. Chem. 273:26273.
    7. Nykjaer, A. et al. (2004) Nature 427:843.
    8. Teng, H.K. et al. (2005) J. Neurosci. 25:5455.
    9. Chen, Z-Y. et al. (2004) J. Neurosci. 25:6156.
    10. Shi, J. and K.V. Kandror (2005) Dev. Cell 9:99.
    11. Nielsen, M.S. et al. (1999) J. Biol. Chem. 274:8832.
  • Entrez Gene IDs:
    6272 (Human); 20661 (Mouse)
  • Alternate Names:
    100 kDa NT receptor; Glycoprotein 95; Gp95; Gp95LDLCQ6; Neurotensin receptor 3; NT3NTR3; Ntr3; SORT1; sortilin 1; Sortilin
Related Research Areas
Citations:

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.

1 Citations: Showing 1 - 1

  1. Role of endogenous brain-derived neurotrophic factor and sortilin in B cell survival.
    Authors: Fauchais AL, Lalloue F, Lise MC, Boumediene A, Preud'homme JL, Vidal E, Jauberteau MO
    J. Immunol., 2008;181(5):3027-38.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
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