Figure 1. Western blot analysis of ZAC/Plagl1 using anti-ZAC/Plagl1 antibody (MBS1755204).
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 30 ug of sample under reducing conditions.
Lane 1: mouse heart tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-ZAC/Plagl1 antigen affinity purified polyclonal antibody (#MBS1755204) at 0.5 ug/mL overnight at 4 degree 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 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for ZAC/Plagl1 at approximately 51 kDa. The expected band size for ZAC/Plagl1 is at 51 kDa.
Figure 2. IF analysis of ZAC/Plagl1 using anti-ZAC/Plagl1 antibody (MBS1755204).
ZAC/Plagl1 was detected in an immunocytochemical section of RM1 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 5 ug/mL rabbit anti-ZAC/Plagl1 Antibody (MBS1755204) overnight at 4 degree C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Figure 3. Flow Cytometry analysis of ANA-1 cells using anti-ZAC/Plagl1 antibody (MBS1755204).
Overlay histogram showing ANA-1 cells stained with MBS1755204 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ZAC/Plagl1 Antibody (MBS1755204, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG (MBS177405, 5-10 ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
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