Overview

  • Product name

    Anti-PC1/3 antibody [EPR21908]
    See all PC1/3 primary antibodies
  • Description

    Rabbit monoclonal [EPR21908] to PC1/3
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, IHC-Fr, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human PC1/3 aa 150-500. The exact sequence is proprietary.
    Database link: P29120

  • Positive control

    • WB: Beta-TC-6 whole cell lysate. Mouse hypothalamus lysate. IHC-P: Human pancreas and colon tissue. Mouse and rat pancreas tissue. IHC-Fr: Mouse and rat pancreas tissue. ICC/IF: Beta-TC-6 cells. Flow Cytometry: Beta-TC-6 cells. IP: Beta-TC-6 cells.
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab220363 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
WB 1/1000. Predicted molecular weight: 84 kDa.
IHC-P 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IHC-Fr 1/500.
ICC/IF 1/100.
Flow Cyt 1/50.
IP 1/30.

Target

  • Function

    Involved in the processing of hormone and other protein precursors at sites comprised of pairs of basic amino acid residues. Substrates include POMC, renin, enkephalin, dynorphin, somatostatin and insulin.
  • Involvement in disease

    Defects in PCSK1 are the cause of proprotein convertase 1 deficiency (PC1 deficiency) [MIM:600955]. PC1 deficiency is characterized by obesity, hypogonadism, hypoadrenalism, reactive hypoglycemia as well as marked small-intestinal absorptive dysfunction It is due to impaired processing of prohormones.
  • Sequence similarities

    Belongs to the peptidase S8 family. Furin subfamily.
  • Cellular localization

    Cytoplasmic vesicle > secretory vesicle. Localized in the secretion granules.
  • Information by UniProt
  • Database links

  • Alternative names

    • BDP antibody
    • NEC 1 antibody
    • NEC1 antibody
    • NEC1_HUMAN antibody
    • Neuroendocrine convertase 1 antibody
    • PC1 antibody
    • PC3 antibody
    • PCSK1 antibody
    • Prohormone convertase 1 antibody
    • Prohormone convertase 1/3 antibody
    • Prohormone convertase 3 antibody
    • Proprotein convertase 1 antibody
    • SPC3 antibody
    see all

Images

  • All lanes : Anti-PC1/3 antibody [EPR21908] (ab220363) at 1/1000 dilution

    Lane 1 : Beta-TC-6 (mouse pancreas insulinoma beta cell) whole cell lysate
    Lane 2 : 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 1/100000 dilution

    Predicted band size: 84 kDa


    Exposure time: 125 seconds


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

    The PC1/3 protein undergoes multiple intracellular cleavage steps to its 66 kDa mature form; the 87 kDa represents the ProPC1/3 intermediate (PMID: 26778167).

    Negative control: NIH/3T3 (PMID: 9405066; PMID:15143067).

  • Immunohistochemical analysis of frozen rat pancreas tissue labeling PC1/3 with ab220363 at 1/500 dilution (green), followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at a 1/1000 dilution. Positive cytoplasmic staining in rat pancreatic islet (PMID: 25976560) is observed. Counter stained with DAPI (blue).
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit used at a 1/1000 dilution.

    Perform heat-mediated antigen retrieval by using Tris-EDTA buffer (10mM Tris base pH 9.0, 1mM EDTA, 0.05% Tween 20).

  • Anti-PC1/3 antibody [EPR21908] (ab220363) at 1/1000 dilution + Mouse hypothalamus lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 84 kDa


    Exposure time: 3 minutes


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

    The 66 kDa band is the mature form of PC1/3 (PMID: 26778167).

  • Immunohistochemical analysis of frozen mouse pancreas tissue labeling PC1/3 with ab220363 at 1/500 dilution (green), followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at a 1/1000 dilution. Positive cytoplasmic staining in mouse pancreatic islet (PMID: 25976560) is observed. Counter stained with DAPI (blue).
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit used at a 1/1000 dilution.

    Perform heat-mediated antigen retrieval by using Tris-EDTA buffer (10mM Tris base pH 9.0, 1mM EDTA, 0.05% Tween 20).

  • Immunohistochemical analysis of paraffin-embedded rat pancreas tissue labeling PC1/3 with ab220363 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining in rat pancreatic islets (PMID: 7925129; PMID: 21190012) is observed. Counter stained with Hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat-mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0).

  • Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue labeling PC1/3 with ab220363 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining in mouse pancreatic islets (PMID: 7925129; PMID: 21190012) is observed. Counter stained with Hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat-mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0).

  • Immunohistochemical analysis of paraffin-embedded human colon tissue labeling PC1/3 with ab220363 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Sporadic cytoplasmic staining (arrows) in human colon (PMID: 18706454) is observed. Counter stained with Hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat-mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0).

  • Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling PC1/3 with ab220363 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining in human pancreatic islets (PMID: 7925129; PMID: 21190012) is observed. Counter stained with Hematoxylin.
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat-mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Beta-TC-6 (mouse pancreas insulinoma beta cell) cells labeling PC1/3 with ab220363 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing cytoplasmic staining in Beta-TC-6 cell line.

    Negative cell control: NIH/3T3 (PMID: 9405066; PMID:15143067).

    Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red). The nuclear counter stain is DAPI (blue). The negative control is the secondary antibody only.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized Beta-TC-6 (mouse pancreas insulinoma beta cell) cell line labeling PC1/3 with ab220363 at 1/50 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) 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 dilution was used as the secondary antibody.

  • PC1/3 was immunoprecipitated from 0.35 mg Beta-TC-6 (mouse pancreas insulinoma beta cell) whole cell lysate with ab220363 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab220363 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
    Lane 1: Beta-TC-6 (mouse pancreas insulinoma beta cell) whole cell lysate 10 µg (Input).
    Lane 2: ab220363 IP in Beta-TC-6 whole cell lysate (+).
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab220363 in Beta-TC-6 whole cell lysate (-).


    Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 3 minutes.

    The PC1/3 protein undergoes multiple intracellular cleavage steps to its 66 kDa mature form (PMID: 26778167).

References

ab220363 has not yet been referenced specifically in any publications.

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