Key Product Details

Species Reactivity

Validated:

Mouse

Cited:

Human, Mouse, Rat

Applications

Validated:

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

Cited:

Immunohistochemistry, Western Blot, Immunocytochemistry, In vivo assay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all Sortilin Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse Sortilin
Gly76-Asn753
Accession # Q6PHU5

Specificity

Detects mouse Sortilin in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 30% cross‑reactivity with recombinant human Sortilin is observed.

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 Mouse Sortilin Antibody

Detection of Mouse Sortilin antibody by Western Blot.

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 antibody in Mouse Brain by Immunohistochemistry (IHC-Fr).

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 antibody by Simple WesternTM.

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.

Detection of Mouse Sortilin by Western Blot

Detection of Mouse Sortilin by Western Blot

Primary cultured Schwann cells express sortilin. (A) Double-labeled immunofluorescence microscopy of primary cultured Schwann cells show sortilin (green) in wild-type (WT) Schwann cells (red; identified by the Schwann cell marker s100 beta ), but not in cells derived from Sort1–/– rat sciatic nerves. Nuclei are labeled with Hoechst (blue). Scale bar 50 μm. (B) Immunoblot analysis of sortilin in lysates of WT or Sort1–/– primary Schwann cells shows sortilin expression in WT. (C) The viability of Sort1–/– Schwann cells is like WT Schwann cells with (95.89 ± 1.55% vs. 94.30 ± 1.77%, p > 0.99) or without NT-3 stimulation (93.8 ± 1.52% vs. 93.29 ± 1.89%, p > 0.99). Viable cells were identified in Schwann cell cultures 24 h after stimulation with or without NT-3 by calcein (green) fluorescence, while dead cells were identified by ethidium homodimer-1 (red) fluorescence. Scale bar 100 μm. Viability reflects mean% calcein-positive cells ± SEM, n = 4. (D) Schwann cell proliferation (percentage of Ki-67% cells) was not significantly different in cultures of Sort1–/– Schwann cells compared to WT Schwann cell cultures with (4.82 ± 1.54% vs. 1.05 ± 0.25%, p = 0.4381) or without NT-3 stimulation (4.09 ± 1.98% vs. 1.37 ± 0.56%, p > 0.99). The proliferation marker, Ki-67 (green), was identified by immunofluorescence in Schwann cell cultures 48 h after stimulation with or without NT-3. Nuclei are labeled with Hoechst (blue). Scale bar 100 μm. Proliferation rates reflect% Ki-67 positive cells ± S.E.M., n = 3. Statistical significance was analyzed using two-way ANOVA with Bonferroni’s multiple comparisons post hoc test. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35634462), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse Sortilin by Western Blot

Detection of Mouse Sortilin by Western Blot

Neurotrophin signaling is decreased in Sort1–/– Schwann cells. (A) pAkt, pMAPK, pRSK, and pCREB were quantified by densitometry of immunoblots from lysates of NT-3 or BDNF stimulated wild-type (WT) or Sort1–/– primary cultures of Schwann cells and normalized against beta -actin. pMAPK, pRSK, and pCREB levels are significantly lower in Sort1–/– Schwann cells compared to WT Schwann cells. Data represent the mean ± SEM for n = 3. (B) The relative wound healing closure time was similar in Sort1–/– Schwann cells relative to WT Schwann cells with or without 10% FBS (positive control) or NT-3. Furthermore, the relative wound healing closure time indicates that NT-3 does not increase the relative wound healing closure time significantly in WT Schwann cells compared to unstimulated controls; 10% FBS significantly increased the relative wound healing closure time at 27–39 h (*p < 0.05). (C)Sort1–/– Schwann cells showed similar myelin formation relative to WT Schwann cells. Co-cultures of DRG neurons and Schwann cells were fixed 11 days after myelin induction with or without NT-3 stimulation. The myelin segments were identified with an antibody against myelin basic protein (MBP; green) and the neurites were identified with antibodies against neurofilament (red). Hoechst identifies nuclei (blue). Statistical significance was determined by two-way ANOVA with Bonferroni’s multiple comparisons post hoc test. Data represents mean ± SEM, n = 3 (*p < 0.05, **p < 0.005, ***p < 0.001, ****p < 0.0001). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35634462), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse Sortilin by Western Blot

Detection of Mouse Sortilin by Western Blot

Primary cultured Schwann cells express sortilin. (A) Double-labeled immunofluorescence microscopy of primary cultured Schwann cells show sortilin (green) in wild-type (WT) Schwann cells (red; identified by the Schwann cell marker s100 beta ), but not in cells derived from Sort1–/– rat sciatic nerves. Nuclei are labeled with Hoechst (blue). Scale bar 50 μm. (B) Immunoblot analysis of sortilin in lysates of WT or Sort1–/– primary Schwann cells shows sortilin expression in WT. (C) The viability of Sort1–/– Schwann cells is like WT Schwann cells with (95.89 ± 1.55% vs. 94.30 ± 1.77%, p > 0.99) or without NT-3 stimulation (93.8 ± 1.52% vs. 93.29 ± 1.89%, p > 0.99). Viable cells were identified in Schwann cell cultures 24 h after stimulation with or without NT-3 by calcein (green) fluorescence, while dead cells were identified by ethidium homodimer-1 (red) fluorescence. Scale bar 100 μm. Viability reflects mean% calcein-positive cells ± SEM, n = 4. (D) Schwann cell proliferation (percentage of Ki-67% cells) was not significantly different in cultures of Sort1–/– Schwann cells compared to WT Schwann cell cultures with (4.82 ± 1.54% vs. 1.05 ± 0.25%, p = 0.4381) or without NT-3 stimulation (4.09 ± 1.98% vs. 1.37 ± 0.56%, p > 0.99). The proliferation marker, Ki-67 (green), was identified by immunofluorescence in Schwann cell cultures 48 h after stimulation with or without NT-3. Nuclei are labeled with Hoechst (blue). Scale bar 100 μm. Proliferation rates reflect% Ki-67 positive cells ± S.E.M., n = 3. Statistical significance was analyzed using two-way ANOVA with Bonferroni’s multiple comparisons post hoc test. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35634462), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse Sortilin by Western Blot

Detection of Mouse Sortilin by Western Blot

Neurotrophin signaling is decreased in Sort1–/– Schwann cells. (A) pAkt, pMAPK, pRSK, and pCREB were quantified by densitometry of immunoblots from lysates of NT-3 or BDNF stimulated wild-type (WT) or Sort1–/– primary cultures of Schwann cells and normalized against beta -actin. pMAPK, pRSK, and pCREB levels are significantly lower in Sort1–/– Schwann cells compared to WT Schwann cells. Data represent the mean ± SEM for n = 3. (B) The relative wound healing closure time was similar in Sort1–/– Schwann cells relative to WT Schwann cells with or without 10% FBS (positive control) or NT-3. Furthermore, the relative wound healing closure time indicates that NT-3 does not increase the relative wound healing closure time significantly in WT Schwann cells compared to unstimulated controls; 10% FBS significantly increased the relative wound healing closure time at 27–39 h (*p < 0.05). (C)Sort1–/– Schwann cells showed similar myelin formation relative to WT Schwann cells. Co-cultures of DRG neurons and Schwann cells were fixed 11 days after myelin induction with or without NT-3 stimulation. The myelin segments were identified with an antibody against myelin basic protein (MBP; green) and the neurites were identified with antibodies against neurofilament (red). Hoechst identifies nuclei (blue). Statistical significance was determined by two-way ANOVA with Bonferroni’s multiple comparisons post hoc test. Data represents mean ± SEM, n = 3 (*p < 0.05, **p < 0.005, ***p < 0.001, ****p < 0.0001). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35634462), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Mouse Sortilin Antibody

Application
Recommended Usage

Blockade of Receptor-ligand Interaction

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.

Immunohistochemistry

1-15 µg/mL
Sample: Perfusion fixed frozen sections of mouse brain (rostral ventral medulla)

Simple Western

10 µg/mL
Sample:   Mouse brain (cortex) tissue

Western Blot

1 µg/mL
Sample: Mouse brain (total) tissue, mouse brain (cerebellum) tissue, and mouse brain (cortex) tissue

Reviewed Applications

Read 8 reviews rated 5 using AF2934 in the following applications:

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). 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.

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

Q6PHU5

Additional Sortilin Products

Product Documents for Mouse Sortilin Antibody

Certificate of Analysis

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Product Specific Notices for Mouse Sortilin Antibody

For research use only

Citations for Mouse Sortilin Antibody

Customer Reviews for Mouse Sortilin Antibody (8)

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8 Customer Ratings
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Customer Images

Showing  1 - 5 of 8 reviews Showing All
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  • Name: Anonymous
    Application: Immunohistochemistry-Frozen
    Sample Tested: Dorsal root ganglia
    Species: Mouse
    Verified Customer | Posted 12/19/2014
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: Primary dorsal root ganglia neurons
    Species: Mouse
    Verified Customer | Posted 12/19/2014
  • Name: Anonymous
    Application: Immunohistochemistry-Frozen
    Sample Tested: Spinal cord cryosections and dorsal root ganglia
    Species: Mouse
    Verified Customer | Posted 12/19/2014
  • Name: Anonymous
    Application: Immunoprecipitation
    Sample Tested: Endogenous mouse sortilin
    Species: Mouse
    Verified Customer | Posted 12/19/2014
  • Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: Primary neurons
    Species: Mouse
    Verified Customer | Posted 12/19/2014
    ICC analysis of Sortilin in a DIV12 hippocampal neuron
    Mouse Sortilin Antibody AF2934
  • Name: Anonymous
    Application: Immunohistochemistry-Frozen
    Sample Tested: Cryosections
    Species: Mouse
    Verified Customer | Posted 12/19/2014
  • Mouse Sortilin Antibody
    Name: Anonymous
    Application: Immunoprecipitation
    Sample Tested: mouse cryosections
    Species: Mouse
    Verified Customer | Posted 12/16/2014
  • Mouse Sortilin Antibody
    Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: mouse cryosections
    Species: Mouse
    Verified Customer | Posted 12/16/2014

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