Anti-Myosin Phosphatase/PPP1R12A Antibody

Figure 1. Western blot analysis of PPP1R12A using anti-PPP1R12A antibody (PA1681).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human Jurka7 whole cell lysates,
Lane 3: human HEK293 whole cell lysates,
Lane 4: monkey COS-7 whole cell lysates,
Lane 5: human Raji whole cell lysates,
Lane 6: human K562 whole cell lysates,
Lane 7: human Caco-2 whole cell lysates,
Lane 8: human HepG2 whole cell lysates,
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-PPP1R12A antigen affinity purified polyclonal antibody (Catalog # PA1681) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for PPP1R12A at approximately 140KD. The expected band size for PPP1R12A is at 115KD.

Figure 2. Flow Cytometry analysis of Hela cells using anti-PPP1R12A antibody (PA1681 ).
Overlay histogram showing Hela cells stained with PA1681 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PPP1R12A Antibody (PA1681 ,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Figure 3. Flow Cytometry analysis of U251 cells using anti-PPP1R12A antibody (PA1681 ).
Overlay histogram showing U251 cells stained with PA1681 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PPP1R12A Antibody (PA1681 ,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Figure 4. Western blot analysis of PPP1R12A using anti-PPP1R12A antibody (PA1681).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: rat brain tissue lysates,
Lane 2: rat C6 whole cell lysates,
Lane 3: mouse liver tissue lysates,
Lane 4: mouse NIH/3T3 whole cell lysates,
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-PPP1R12A antigen affinity purified polyclonal antibody (Catalog # PA1681) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for PPP1R12A at approximately 140KD. The expected band size for PPP1R12A is at 115KD.

Figure 5. IF analysis of PPP1R12A using anti-PPP1R12A antibody (PA1681).
PPP1R12A was detected in immunocytochemical section of SiHa cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-PPP1R12A Antibody (PA1681) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.