Anti foxl2 antibody

Ovaries were fixed with 4% PFA (paraformaldehyde) at 4°C overnight, which was then graded to 30% sucrose, and ovaries were then embedded in O.C.T compound (Sakura Fine tek). Each sample was sliced at 6-μm thickness. After removing the O.C.T compound, slides were incubated with 3% skim milk in PBST (PBS, 0.1% Tween-20) for 1 hour. Primary antibody reactions were performed with the following dilutions (Anti-DAZL antibody, Abcam, 1:200 / Anti-FOXL2 antibody, Abcam, 1:200) at 4°C overnight. After washing with PBST, secondary antibody reaction was performed with the following dilutions (Alexa 488 Donkey anti-Rabbit, Life technologies, 1:400 /Alexa 594 Donkey anti-Goat, Life technologies, 1:400) at RT for 90 min. Slides then were counter-stained by DAPI at RT for 15 min. Fluorescent images were obtained by confocal microscopy FV1200 (Olympus).

Ovaries were fixed in 4% PFA (paraformaldehyde) at 4°C overnight and embedded in paraffin wax. Each sample was sliced at 6-μm thickness and placed on glass slides. After removing the paraffin wax and autoclaving in antigen unmasking solution/high pH (Vector Laboratories), glass slides were washed in PBST (PBS, 0.1%Tween-20) and pre-incubated in 3% skim milk in PBST blocking solution at RT for 1 hour. The slides were reacted with primary antibodies (Anti-DAZL antibody, Abcam, 1:200 / Anti-FOXL2 antibody, Abcam, 1:200/ Anti-FLAG antibody, SIGMA, 1:10000) at 4°C overnight. Then, slides were washed with PBST and incubated with second antibodies (Alexa 488 Donkey anti-Rabbit, Life technologies, 1:1000 /Alexa 594 Donkey anti-Mouse, Life technologies, 1:1000/Alexa 594 Donkey anti-Goat, Life technologies, 1:1000 / Cy5 Donkey anti-goat, Rockland, 1:1000) at RT for 60 min. DNA was counter-stained with DAPI, and fluorescent images were obtained using confocal microscopy FV1200 (Olympus).