Protein isolation in zebrafish - (Feb/27/2011 )
I have no experience in protein work, but I need to isolate protein from 24 - 72 hpf zebrafish embryos. I've just looked through the Zebrafish Book on Zfin and there are two protocols there for protein isolation and both are very different from each other. One protocol pretty much just requires the homogenisation of the embryos in the SDS sample buffer and the other requires deyolking the embryos before proceeding. What is the purpose of deyolking the embyros and if anyone has a standard protocol they've tried and works could you please share?
De-yolking ensures that the proteins are from the embryo, not the yolk, so you are more likely to see if non-maternal genes are being expressed. The protocols on Zfin work just fine.