Rat anti tubulin antibody

Progression of cells through meiosis in each timecourse included here was determined by quantification of nuclear morphology by DAPI staining (Vectashield, Vector) of ethanol-permeabilized cells adhered to a polylysine-treated glass slide. Prior to anaphase I, cells show mononucleate morphology, at and after anaphase I and before anaphase II, cells show binucleate morphology, during and following anaphase II, cells show tetranucleate morphology. All timecourses were also assessed at 24 hours after transfer to sporulation media by brightfield microscopy to ensure high efficiency of spore formation, which we typically observe at ~90%. The Ndt80 induction experiment (Fig. 5C–I, S5) was also staged using indirect immunofluorescence of alpha-tubulin, using a rat anti-tubulin antibody (Serotec) at a dilution of 1:200 and anti-rat FITC antibody (Jackson ImmunoResearch Laboratories) at 1:100. Fluorescent microscopy was done on a DeltaVision microscope with a 100X objective.