Recombinant
RabMAb

Recombinant Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)

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Overview

  • Product name

    Anti-Sortilin/NT3 antibody [EPR23093-58]
    See all Sortilin/NT3 primary antibodies

  • Description

    Rabbit monoclonal [EPR23093-58] to Sortilin/NT3

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt, ICC/IF, WB, IHC-Pmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human Sortilin/NT3 aa 50-800. The exact sequence is proprietary.
    Database link: Q99523

  • Positive control

    • WB: Human brain, SH-SY5Y, mouse brain, mouse lung, rat brain, rat lung, Neuro-2a, C6, RAW 264.7, PC-12 and NIH/3T3 lysates. IHC-P: Human endometrium, human ovarian carcinoma, breast carcinoma and the paired adjacent breast tissue. Mouse and rat cerebrum tissue. ICC/IF: SH-SY5Y and Neuro-2a cells. FC: SH-SY5Y and Neuro-2a cells.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

Our Abpromise guarantee covers the use of ab263864 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt 1/600.

 

 
ICC/IF 1/100.

 

 
WB 1/1000. Predicted molecular weight: 92 kDa.

 

 
IHC-P 1/5000.

 

 
  • Application notes
    Is unsuitable for IP.

    • Target

    • Function

      Functions as a sorting receptor in the Golgi compartment and as a clearance receptor on the cell surface. Required for protein transport from the Golgi apparatus to the lysosomes by a pathway that is independent of the mannose-6-phosphate receptor (M6PR). Also required for protein transport from the Golgi apparatus to the endosomes. Promotes neuronal apoptosis by mediating endocytosis of the proapoptotic precursor forms of BDNF (proBDNF) and NGFB (proNGFB). Also acts as a receptor for neurotensin. May promote mineralization of the extracellular matrix during osteogenic differentiation by scavenging extracellular LPL. Probably required in adipocytes for the formation of specialized storage vesicles containing the glucose transporter SLC2A4/GLUT4 (GLUT4 storage vesicles, or GSVs). These vesicles provide a stable pool of SLC2A4 and confer increased responsiveness to insulin. May also mediate transport from the endoplasmic reticulum to the Golgi.

    • Tissue specificity

      Expressed at high levels in brain, spinal cord, heart, skeletal muscle, thyroid, placenta and testis. Expressed at lower levels in lymphoid organs, kidney, colon and liver.

    • Involvement in disease

      Note=A common polymorphism located in a non-coding region between CELSR2 and PSRC1 alters a CEBP transcription factor binding site and is responsible for changes in hepatic expression of SORT1. Altered SORT1 expression in liver affects low density lipoprotein cholesterol levels in plasma and is associated with susceptibility to myocardial infarction.

    • Sequence similarities

      Belongs to the VPS10-related sortilin family. SORT1 subfamily.
      Contains 9 BNR repeats.

    • Domain

      The N-terminal propeptide may facilitate precursor transport within the Golgi stack. Intrachain binding of the N-terminal propeptide and the extracellular domain may also inhibit premature ligand binding.
      The extracellular domain may be shed following protease cleavage in some cell types.

    • Post-translational
      modifications

      The N-terminal propeptide is cleaved by furin and possibly other homologous proteases.

    • Cellular localization

      Membrane. Endoplasmic reticulum membrane. Endosome membrane. Golgi apparatus > Golgi stack membrane. Lysosome membrane. Nucleus membrane. Cell membrane. Lysosome membrane. Localized to membranes of the endoplasmic reticulum, endosomes, Golgi stack, lysosomes and nucleus. A small fraction of the protein is also localized to the plasma membrane. May also be found in SLC2A4/GLUT4 storage vesicles (GSVs) in adipocytes. Localization to the plasma membrane in adipocytes may be enhanced by insulin.
    • Information by UniProt

    • Database links

      see all

    • Alternative names

      • 100 kDa NT receptor antibody
      • Glycoprotein 95 antibody
      • Gp 95 antibody
      • Gp95 antibody
      • LDLCQ6 antibody
      • Neurotensin receptor 3 antibody
      • NT 3 antibody
      • NT3 antibody
      • NTR 3 antibody
      • NTR3 antibody
      • OTTHUMP00000013784 antibody
      • SORT 1 antibody
      • SORT_HUMAN antibody
      • SORT1 (gene name) antibody
      • Sort1 antibody
      • Sortilin 1 antibody
      • Sortilin antibody
      see all

    Images

    • Western blot - Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)
      Western blot - Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)
      All lanes : Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864) at 1/1000 dilution

      Lane 1 : C6 (rat glial tumor glial cell) whole cell lysate
      Lane 2 : RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
      Lane 3 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
      Lane 4 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

      Predicted band size: 92 kDa
      Observed band size: 100 kDa
      why is the actual band size different from the predicted?



      Blocking and diluting buffer and concentration: 5% NFDM/TBST.

      Exposure time: 48 seconds.

    • Western blot - Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)
      Western blot - Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)
      All lanes : Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864) at 1/1000 dilution

      Lane 1 : Human brain lysate
      Lane 2 : SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate
      Lane 3 : Mouse brain lysate
      Lane 4 : Mouse lung lysate
      Lane 5 : Rat brain lysate
      Lane 6 : Rat lung lysate
      Lane 7 : Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Lane 1 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
      Lanes 2-7 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

      Predicted band size: 92 kDa
      Observed band size: 100 kDa why is the actual band size different from the predicted?



      Blocking and diluting buffer and concentration: 5% NFDM/TBST.

      Exposure time: Lanes 1-2: 48 seconds; Lanes 3-7: 6 seconds.

    • Flow Cytometry - Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)
      Flow Cytometry - Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)

      Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Neuro-2a (Mouse neuroblastoma neuroblast) cells labeling Sortilin/NT3 with ab263864 at 1/600 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1\2000 was used as the secondary antibody.

    • Flow Cytometry - Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)
      Flow Cytometry - Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)

      Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling Sortilin/NT3 with ab263864 at 1/600 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1\2000 was used as the secondary antibody.

    • Immunocytochemistry/ Immunofluorescence - Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)
      Immunocytochemistry/ Immunofluorescence - Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)

      Immunofluorescent analysis of 100% methanol-fixed Neuro-2a (Mouse neuroblastoma neuroblast) cells labeling Sortilin/NT3 with ab263864 at 1\100 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1\1000 dilution (Green). Confocal image showing golgi staining in Neuro-2a cell line. 100% methanol fixation is recommended. Counterstained with ab169276 Anti-GM130 antibody-Golgi Marker (1/500 dilution) and ab150120 AlexaFluor®594 Goat anti-Mouse secondary (1/1000 dilution, red). The nuclear counterstain was DAPI (Blue).

      The negative controls are as follows:

      -ve control 1: ab263864 at 1/100 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/500 dilution.

      -ve control 2: ab169276 Anti-GM130 antibody- Golgi Marker at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution.

    • Immunocytochemistry/ Immunofluorescence - Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)
      Immunocytochemistry/ Immunofluorescence - Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)

      Immunofluorescent analysis of 100% methanol-fixed SH-SY5Y (Human neuroblastoma epithelial cell) cells labelling Sortilin/NT3 with ab263864 at 1\100 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1\1000 dilution (Green). Confocal image showing cytoplasmic staining in SH-SY5Y cell line. 100% methanol fixation is recommended. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

      Secondary antibody only control: Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1\1000 dilution.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)

      Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling Sortilin/NT3 with ab263864 at 1/5000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on rat cerebrum is observed. The section was incubated with ab263864 for 10 mins at RT. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)

      Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling Sortilin/NT3 with ab263864 at 1/5000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on mouse cerebrum iso bserved. The section was incubated with ab263864 for 10 mins at RT. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)

      Immunohistochemical analysis of paraffin-embedded human breast carcinoma and the paired adjacent breast tissue tissue labeling Sortilin/NT3 with ab263864 at 1/5000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human breast carcinoma (panel A) and weak staining on the paired adjacent breast tissue (panel B) (PMID: 25871389) is observed. The section was incubated with ab263864 for 10 mins at RT. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)

      Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma tissue labeling Sortilin/NT3 with ab263864 at 1/5000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human ovarian carcinoma (PMID: 23407681) is observed. The section was incubated with ab263864 for 10 mins at RT. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sortilin/NT3 antibody [EPR23093-58] (ab263864)

      Immunohistochemical analysis of paraffin-embedded human endometrium tissue labeling Sortilin/NT3 with ab263864 at 1/5000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human endometrium (PMID: 25662808) is observed. The section was incubated with ab263864 for 10 mins at RT. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References

    Publishing research using ab263864? Please let us know so that we can cite the reference in this datasheet.

    ab263864 has not yet been referenced specifically in any publications.

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