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RecombinantRabMAb

Recombinant Anti-PC1/3 antibody [EPR21908] (ab220363)

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Western blot - Anti-PC1/3 antibody [EPR21908] (ab220363)
  • Immunohistochemistry (Frozen sections) - Anti-PC1/3 antibody [EPR21908] (ab220363)
  • Western blot - Anti-PC1/3 antibody [EPR21908] (ab220363)
  • Immunohistochemistry (Frozen sections) - Anti-PC1/3 antibody [EPR21908] (ab220363)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PC1/3 antibody [EPR21908] (ab220363)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PC1/3 antibody [EPR21908] (ab220363)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PC1/3 antibody [EPR21908] (ab220363)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PC1/3 antibody [EPR21908] (ab220363)
  • Immunocytochemistry/ Immunofluorescence - Anti-PC1/3 antibody [EPR21908] (ab220363)
  • Flow Cytometry (Intracellular) - Anti-PC1/3 antibody [EPR21908] (ab220363)
  • Immunoprecipitation - Anti-PC1/3 antibody [EPR21908] (ab220363)
  • Anti-PC1/3 antibody [EPR21908] (ab220363)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21908] to PC1/3
  • Suitable for: WB, IHC-P, IHC-Fr, ICC/IF, IP, Flow Cyt (Intra)
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 647 Carrier Free

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, IP, Flow Cyt (Intra)more details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • 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 Cyt (intra): Beta-TC-6 cells. IP: Beta-TC-6 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

The Abpromise guarantee

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.
IP
1/30.
Flow Cyt (Intra)
1/50.
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.
IP
1/30.
Flow Cyt (Intra)
1/50.

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

    see all

  • 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

  • Western blot - Anti-PC1/3 antibody [EPR21908] (ab220363)
    Western blot - Anti-PC1/3 antibody [EPR21908] (ab220363)
    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

    Developed using the ECL technique.

    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).

    This blot was developed using a higher sensitivity ECL substrate.

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

  • Immunohistochemistry (Frozen sections) - Anti-PC1/3 antibody [EPR21908] (ab220363)
    Immunohistochemistry (Frozen sections) - Anti-PC1/3 antibody [EPR21908] (ab220363)

    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).

  • Western blot - Anti-PC1/3 antibody [EPR21908] (ab220363)
    Western blot - Anti-PC1/3 antibody [EPR21908] (ab220363)
    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

    Developed using the ECL technique.

    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).

    This blot was developed using a higher sensitivity ECL substrate.

  • Immunohistochemistry (Frozen sections) - Anti-PC1/3 antibody [EPR21908] (ab220363)
    Immunohistochemistry (Frozen sections) - Anti-PC1/3 antibody [EPR21908] (ab220363)

    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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PC1/3 antibody [EPR21908] (ab220363)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PC1/3 antibody [EPR21908] (ab220363)

    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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PC1/3 antibody [EPR21908] (ab220363)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PC1/3 antibody [EPR21908] (ab220363)

    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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PC1/3 antibody [EPR21908] (ab220363)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PC1/3 antibody [EPR21908] (ab220363)

    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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PC1/3 antibody [EPR21908] (ab220363)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PC1/3 antibody [EPR21908] (ab220363)

    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).

  • Immunocytochemistry/ Immunofluorescence - Anti-PC1/3 antibody [EPR21908] (ab220363)
    Immunocytochemistry/ Immunofluorescence - Anti-PC1/3 antibody [EPR21908] (ab220363)

    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 Cytometry (Intracellular) - Anti-PC1/3 antibody [EPR21908] (ab220363)
    Flow Cytometry (Intracellular) - Anti-PC1/3 antibody [EPR21908] (ab220363)

    Intracellular 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. 

     

     

  • Immunoprecipitation - Anti-PC1/3 antibody [EPR21908] (ab220363)
    Immunoprecipitation - Anti-PC1/3 antibody [EPR21908] (ab220363)

    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).

  • Anti-PC1/3 antibody [EPR21908] (ab220363)
    Anti-PC1/3 antibody [EPR21908] (ab220363)

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 (2)

Publishing research using ab220363? Please let us know so that we can cite the reference in this datasheet.

ab220363 has been referenced in 2 publications.

  • Ma L  et al. Sennoside A Induces GLP-1 Secretion Through Activation of the ERK1/2 Pathway in L-Cells. Diabetes Metab Syndr Obes 13:1407-1415 (2020). PubMed: 32425572
  • Kreutzberger AJB  et al. Distinct insulin granule subpopulations implicated in the secretory pathology of diabetes types 1 and 2. Elife 9:N/A (2020). PubMed: 33164744

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