IF analysis of ZIP Kinase/Dapk3 using anti-ZIP Kinase/Dapk3 antibody (A03300-3). ZIP Kinase/Dapk3 was detected in an immunocytochemical section of RM-1 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 µg/mL rabbit anti-ZIP Kinase/Dapk3 Antibody (A03300-3) 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.

Western blot analysis of ZIP Kinase/Dapk3 using anti-ZIP Kinase/Dapk3 antibody (A03300-3). 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 lung tissue lysates, Lane 2: mouse stomach tissue lysates, Lane 3: rat kidney tissue lysates, Lane 4: rat PC-12 whole cell 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-ZIP Kinase/Dapk3 antigen affinity purified polyclonal antibody (Catalog # A03300-3) 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 1:5000 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 ZIP Kinase/Dapk3 at approximately 53 kDa. The expected band size for ZIP Kinase/Dapk3 is at 53 kDa.