Subcellular localization of cytoplasmic lattice-associated proteins is dependent upon fixation and processing procedures.

ABSTRACT: We and others have recently demonstrated by immuno-EM and mutation analysis that two oocyte-restricted maternal effect genes, PADI6 and MATER, localize, in part, to the oocyte cytoplasmic lattices (CPLs). During these ongoing studies, however, we found that the localization of these factors by confocal immunofluorescence (IF) analysis can vary dramatically depending upon how the oocytes and embryos are processed, with the localization pattern sometimes appearing more uniformly cytoplasmic while at other times appearing to be primarily cortical. We set out to better understand this differential staining pattern by testing a range of IF protocol parameters, changing mainly time and temperature conditions of the primary antibody solution incubation, as well as fixation methods. We found by co

SUBMITTER: Morency E 

PROVIDER: S-EPMC3040232 | biostudies-literature | 2011 Feb

REPOSITORIES: biostudies-literature