Ab84892

Cells on coverslips were fixed in 4% paraformaldehyde in PBS for 10 min followed by one wash with PBS, permeabilization in 0.2% Triton X-100 in PBS for 5 min, and three washes with PBS (5 min each wash). Coverslips were blocked with blocking buffer (10% horse serum, 1% BSA, 0.02% NaN₃, 1× PBS) for 1 h, washed with PBS and incubated with anti-ALMS1 (ab84892, abcam) and anti-acetylated tubulin (T7451, Sigma-Aldrich) in 1% BSA in PBS overnight at 4°C. After washing with PBS, the cells were incubated with 1:1000 dilution of Alexa Fluor® 488 goat anti-mouse IgG (A11001, Invitrogen) and Alexa Fluor® 555 goat anti-rabbit IgG (A21430, invitrogen) for 45 min at room temperature in the dark, washed with PBS, mounted on glass slides using the ProLong Gold Antifade Reagent with DAPI (P36931, invitrogen) and inspected with a Zeiss LSM510 Meta confocal laser scanning microscope. For detecting DNA damage, anti-γH2AX antibody (05-636, Millipore) was used according to the above method except that TBS [50 mM Tris-HCl (pH7.4), 150 mM NaCl] instead of PBS was used.