Anti-Pin1 antibody [EPR18546-317]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Pin1 antibody [EPR18546-317] (AB192036)

Immunofluorescent analysis of 4% PFA-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling Pin1 with ab192036 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 and nuclear staining on HeLa cells.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Pin1 antibody [EPR18546-317] (AB192036)

Intracellular flow cytometric analysis of 4% PFA-fixed, 90% methanol permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling Pin1with ab192036 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Pin1 antibody [EPR18546-317] (AB192036)

Immunofluorescent analysis of 4% PFA-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryonic fibroblast cell line) cells labeling Pin1 with ab192036 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 and nuclear staining on NIH/3T3 cells.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• IP

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Immunoprecipitation - Anti-Pin1 antibody [EPR18546-317] (AB192036)

Pin1 was immunoprecipitated from 0.35 mg of NIH/3T3 (mouse embryonic fibroblast cell line) lysate with ab192036 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab192036 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : NIH/3T3 whole cell lysate 10 μg (Input).

Lane 2 : ab192036 IP in NIH/3T3 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab192036 in NIH/3T3 whole cell lysate.

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

Exposure time : 2 seconds.

All lanes:

Immunoprecipitation - Anti-Pin1 antibody [EPR18546-317] (ab192036)

Predicted band size: 18 kDa

Observed band size: 18 kDa

false

• WB

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Western blot - Anti-Pin1 antibody [EPR18546-317] (AB192036)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure times : Lanes 1-3 : 15 seconds; Lanes 4-6 : 30 seconds.

All lanes:

Western blot - Anti-Pin1 antibody [EPR18546-317] (ab192036) at 1/5000 dilution

Lane 1:

HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate at 20 µg

Lane 2:

MCF7 (human breast adenocarcinoma cell line) cell lysate at 20 µg

Lane 3:

C6 (rat glial tumor cell line) cell lysate at 20 µg

Lane 4:

NIH/3T3 (mouse embryonic fibroblast cell line) cell lysate at 20 µg

Lane 5:

Mouse brain lysate at 20 µg

Lane 6:

Rat brain lysate at 20 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 1/100000 dilution

Predicted band size: 18 kDa

Observed band size: 18 kDa

true

• WB

Supplier Data

Western blot - Anti-Pin1 antibody [EPR18546-317] (AB192036)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Pin1 antibody [EPR18546-317] (ab192036) at 1/2000 dilution

Lane 1:

Human fetal brain lysate at 10 µg

Lane 2:

Human fetal heart lysate at 10 µg

Lane 3:

Human fetal kidney lysate at 10 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/4000 dilution

Predicted band size: 18 kDa,30 kDa

Observed band size: 18 kDa

true

Exposure time: 5s

• WB

Supplier Data

Western blot - Anti-Pin1 antibody [EPR18546-317] (AB192036)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure times : Lanes 1-3 : 5 seconds; Lanes 4-6 : 15 seconds.

All lanes:

Western blot - Anti-Pin1 antibody [EPR18546-317] (ab192036) at 1/2000 dilution

Lane 1:

Mouse heart lysate at 10 µg

Lane 2:

Mouse kidney lysate at 10 µg

Lane 3:

Mouse spleen lysate at 10 µg

Lane 4:

Rat heart lysate at 10 µg

Lane 5:

Rat kidney lysate at 10 µg

Lane 6:

Rat spleen lysate at 10 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 1/100000 dilution

Predicted band size: 18 kDa

Observed band size: 18 kDa

true