Anti-PPP1R1A antibody [EPR28192-90]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-PPP1R1A antibody [EPR28192-90] (AB316931)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage, Left) / Beta-TC-6 (mouse pancreas insulinoma beta cell, Right) cells labelling PPP1R1A with ab316931 at 1/500 dilution (0.1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Negative control : RAW 264.7

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PPP1R1A antibody [EPR28192-90] (AB316931)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling PPP1R1A with ab316931 at 1/2000 (0.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse liver. The section was incubated with ab316931 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PPP1R1A antibody [EPR28192-90] (AB316931)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling PPP1R1A with ab316931 at 1/2000 (0.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse spleen. The section was incubated with ab316931 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PPP1R1A antibody [EPR28192-90] (AB316931)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling PPP1R1A with ab316931 at 1/2000 (0.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on distal convoluted tubule in mouse kidney (PMID : 24231659). The section was incubated with ab316931 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PPP1R1A antibody [EPR28192-90] (AB316931)

Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue labeling PPP1R1A with ab316931 at 1/2000 (0.25 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on islet of mouse pancreas (PMID : 23557701). The section was incubated with ab316931 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PPP1R1A antibody [EPR28192-90] (AB316931)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Beta-TC-6 (mouse pancreas insulinoma beta cell) cells labellingPPP1R1A with ab316931 at 1/500 (1.0 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution

Confocal image showing cytoplasmic with nuclear staining in Beta-TC-6 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Negative control : RAW 264.7 (PMID : 29444892).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution. The Nuclear counterstain was DAPI

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• IP

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Immunoprecipitation - Anti-PPP1R1A antibody [EPR28192-90] (AB316931)

PPP1R1A was immunoprecipitated from 0.35 mg Beta-TC-6 (mouse pancreas insulinoma beta cell) whole cell lysate with ab316931 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab316931 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Beta-TC-6 (mouse pancreas insulinoma beta cell) whole cell lysate
Lane 2 : ab316931 IP in Beta-TC-6 (mouse pancreas insulinoma beta cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab316931 in Beta-TC-6 whole cell lysate

All lanes:

Immunoprecipitation - Anti-PPP1R1A antibody [EPR28192-90] (ab316931) at 1/30 dilution

All lanes:

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

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 67s

• WB

Lab

Western blot - Anti-PPP1R1A antibody [EPR28192-90] (AB316931)

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

Negative control : RAW 264.7, MEF (PMID : 29444892).

ab181602 was used as a GAPDH loading control at 1/200000 dilution

All lanes:

Western blot - Anti-PPP1R1A antibody [EPR28192-90] (ab316931) at 1/1000 dilution

Lane 1:

Beta-TC-6 (mouse pancreas insulinoma beta cell) whole cell lysate at 40 µg

Lane 2:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 40 µg

Lane 3:

MEF (mouse embryo fibroblast) whole cell lysate at 40 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 27 kDa

false

Exposure time: 37s