Anti-Zebrafish si:dkeyp-75b4.10/eslec Antibody

Western blot analysis of Si:dkeyp-75b4.10 using anti-Si:dkeyp-75b4.10 antibody (DZ41383).
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: Marker,
Lane 2: Zebrafish,
Lane 3: Zebrafish body 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-Si:dkeyp-75b4.10 antigen affinity purified polyclonal antibody (DZ41383) 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 Si:dkeyp-75b4.10at approximately 18 kDa. The expected band size for Si:dkeyp-75b4.10 is at 18 kDa.

IF analysis of zebrafish eosinophils using Anti-Zebrafish Si:Dkeyp-75b4.10 Antibody (DZ41383).
Si:Dkeyp-75b4.10 was detected in zebrafish eosinophils in FFPE tissue sections and fixed cells. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 3% goat serum in PBS. The tissue section was then incubated with 4 μg/mL rabbit anti-GFAP antibody (MA1045) and anti-MBP antibody (PA1050) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126), Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:1000 dilution and incubated for 1 hour at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.