passages: PMC-2171703-sec-06__text
passages: sec
passages: Immunocytochemistry
  C2C12 cells grown in 8-chamber slides (Nunc) were fixed in 4% PFA/PBS for 10 min and permeabilized in 0.1% Triton X-100 for 20 min. In some cases, they were fixed and permeabilized in methanol at -20degreesC for 2 min. The fixed and permeabilized cells were blocked in PBS containing 3% BSA (Jackson ImmunoResearch Laboratories) and incubated overnight at 4degreesC with primary antibodies in blocking solution. Immunoreactivity was detected with either Alexa-conjugated secondary antibody (Molecular Probes) or a combination of biotin-conjugated secondary antibody (Vector Laboratories) and Alexa-conjugated streptavidin (Molecular Probes).
  