Mouse monoclonal anti γ tubulin clone gtu 88 t5326

hPSCs embedded in Matrigel were fixed with either 4% PFA at room temperature or ice-cold methanol at -20°C for 15 min. The samples were permeabilized with 0.3% Tx100 – 0.1 M glycine in PBS for 20 min. The primary antibodies mouse monoclonal anti-Oct3/4 (as above, 1:200), rabbit polyclonal anti-aPKC (as above, 1:200), rabbit polyclonal anti-PARD6B (as above, 1:200), mouse monoclonal anti-GM130 (clone 35/GM130; 610822; BD Biosciences; San Jose, US; 1:200), mouse monoclonal anti–γ–tubulin (clone GTU-88; T5326; Sigma, St.Louis, US; 1:500) and mouse monoclonal anti-PODXL (clone 222328; MAB1658; R&D Systems; Minneapolis, US; 1:200) were diluted in blocking buffer (1% Bovine Serum Albumin – A3311; Sigma-Aldrich; St. Louis, US - and 0.1% Tween) and incubated at 4°C over night. Samples were washed three times with PBS-0.1% Tween and incubated with AlexaFluor®-conjugated secondary antibodies (as above).