重组Anti-PC1/3抗体[EPR21908]

• IHC-P

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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).

• IHC-P

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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).

• ICC/IF

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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.

• IHC-P

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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).

• IHC-Fr

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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).

• Flow Cyt (Intra)

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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.

• IHC-P

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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).

• IHC-Fr

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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).

• IP

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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).

All lanes:

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

Predicted band size: 84 kDa

false

• WB

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Western blot - Anti-PC1/3 antibody [EPR21908] (AB220363)

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).

All lanes:

Western blot - Anti-PC1/3 antibody [EPR21908] (ab220363) at 1/1000 dilution

Lane 1:

Beta-TC-6 (mouse pancreas insulinoma beta cell) whole cell lysate at 10 µg

Lane 2:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 84 kDa

true

Exposure time: 125s

• WB

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Western blot - Anti-PC1/3 antibody [EPR21908] (AB220363)

Blocking and Diluting Buffer : 5% NFDM /TBST 87 kDa ProPC1/3; 66 kDa mature form (PMID : 26778167) Lane1 Expsoure Time : 20 seconds Lane2 and Lane3 were developed using a high sensitivity ECL substrate.

All lanes:

Western blot - Anti-PC1/3 antibody [EPR21908] (ab220363) at 1/1000 dilution

Lane 1:

Beta-TC-6 (mouse pancreas insulinoma beta cell) whole cell lysate at 20 µg

Lane 2:

Mouse pancreas lysate at 20 µg

Lane 3:

Mouse brain lysate at 20 µg

Secondary

All lanes:

Western blot - PE Anti-ENPP3/B10 antibody [NP4D6] (<a href='/products/primary-antibodies/pe-enpp3-b10-antibody-np4d6-ab90751'>ab90751</a>) at 1/20000 dilution

Predicted band size: 87 kDa

Observed band size: 66 kDa

false

Exposure time: 80s

• WB

Supplier Data

Western blot - Anti-PC1/3 antibody [EPR21908] (AB220363)

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.

All lanes:

Western blot - Anti-PC1/3 antibody [EPR21908] (ab220363) at 1/1000 dilution

All lanes:

Mouse hypothalamus lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 84 kDa

true

Exposure time: 3min