Human Sortilin Antibody

Catalog # Availability Size / Price Qty
AF3154
AF3154-SP
Detection of Human Sortilin by Western Blot.
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Product Details
Citations (4)
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Human Sortilin Antibody Summary

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

Western Blot Detection of Human Sortilin by Western Blot. View Larger

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.

Flow Cytometry Detection  of Sortilin in Human Blood Monocytes by Flow Cytometry. View Larger

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.

Flow Cytometry Detection  of Sortilin in K562 Human Cell Line by Flow Cytometry. View Larger

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. View Larger

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.

Simple Western Detection  of Human Sortilin by Simple WesternTM. View Larger

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.        

Reconstitution Calculator

Reconstitution Calculator

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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: 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

Product Datasheets

Citations for Human Sortilin Antibody

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.

4 Citations: Showing 1 - 4
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  1. Lack of human-like extracellular sortilin neuropathology in transgenic Alzheimer's disease model mice and macaques
    Authors: FQ Zhou, J Jiang, CM Griffith, PR Patrylo, H Cai, Y Chu, XX Yan
    Alzheimers Res Ther, 2018;10(1):40.
    Species: Primate
    Sample Types: Whole Tissue
    Applications: IHC
  2. Transcriptional control of intestinal cholesterol absorption, adipose energy expenditure and lipid handling by Sortilin
    Authors: S Hagita, MA Rogers, T Pham, JR Wen, AK Mlynarchik, M Aikawa, E Aikawa
    Sci Rep, 2018;8(1):9006.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: WB
  3. proBDNF is modified by advanced glycation end products in Alzheimer's disease and causes neuronal apoptosis by inducing p75 neurotrophin receptor processing
    Authors: C Fleitas, G Piñol-Ripo, P Marfull, D Rocandio, I Ferrer, C Rampon, J Egea, C Espinet
    Mol Brain, 2018;11(1):68.
    Species: Human
    Sample Types: Tissue Section
    Applications: IHC-P
  4. 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 Types: Cell Lysates
    Applications: WB

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