Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Primate, Transgenic Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, Blockade of Receptor-ligand Interaction, Flow Cytometry, Simple Western

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Neutralization, Immunocytochemistry, Bioassay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all Sortilin Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human Sortilin
Ser78-Asn755
Accession # Q99523

Specificity

Detects human and mouse Sortilin in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human Sortilin Antibody

Detection of Human Sortilin antibody by Western Blot.

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 antibody in Human Blood Monocytes antibody by Flow Cytometry.

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 antibody in K562 Human Cell Line antibody by Flow Cytometry.

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.

Sortilin antibody in Human Brain by Immunohistochemistry (IHC-P).

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 antibody by Simple WesternTM.

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.

Applications for Human Sortilin Antibody

Application
Recommended Usage

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.

Flow Cytometry

0.25 µg/106 cells
Sample: Human peripheral blood monocytes and K562 human chronic myelogenous leukemia cell line

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human brain (cerebellum and cortex)

Simple Western

10 µg/mL
Sample: Human brain (cerebellum) tissue

Western Blot

2 µg/mL
Sample: Human motor cortex

Reviewed Applications

Read 2 reviews rated 4.5 using AF3154 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

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  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

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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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.

Alternate Names

Gp95, Ntr3, SORT1

Entrez Gene IDs

6272 (Human); 20661 (Mouse)

Gene Symbol

SORT1

UniProt

Q99523

Additional Sortilin Products

Product Documents for Human Sortilin Antibody

Certificate of Analysis

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Product Specific Notices for Human Sortilin Antibody

For research use only

Citations for Human Sortilin Antibody

Customer Reviews for Human Sortilin Antibody (2)

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  • Human Sortilin Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: Serum
    Species: Human
    Verified Customer | Posted 04/05/2022
    Human Sortilin Antibody AF3154
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: See PMID 22579764
    Species: Other
    Verified Customer | Posted 01/07/2015

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