Protocols
Depolymerization of non-kinetochore microtubules by cold treatment
Incubated cells 10 min on ice in L-15 medium (Invitrogen) supplemented with 20 mM HEPES, pH 7.3. Adjust the pH of the medium on ice, not at room temperature, before use. Fix cells for 10 min at room temperature with 3.7% formaldehyde in 100 mM PIPES at pH 6.8, 10 mM EGTA, 1 mM MgCl2, 0.2% Triton X-100. Process cells for immunofluorescence using standard protocols.