I recommend using PLA (Proximity Ligation Asssay) to detect caspase 3. You must have two different antibodies from different species for example one is rabbit while the other is a mouse that recognize different epitopes .. You can find more info in the PLA protocol .. see Olikns website. The advantage of PLA is that you can see where the protein is localized in intact cells and even relocalization after treatment.

Hi michal! Are you talking about oocytes in GV, GVBD or MII?, Im used to work with eggs in when they are still enclosed by the follicular cells and Caspase3 seems not to have expression in the cytoplasm of the eggs, this is also supported by several papers, however, in fully grown oocyte caspase3 might be present.

Are you gonna do full-mounting? Or life cell imaging?

I can send you some papers later if you want

Hi Randy. It will be oocytes after maturation during their in vitro aging. It will be after fixation for classical immunodetection. Because i would like to use it in porcine oocytes.

Thank you for your offer, it will be helpfull when you recommend me any good paper.

thanks