Immunocytochemistry analysis of Ramos cells labeling Bik with Rabbit anti-Bik antibody (HA751413) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Bik antibody (HA751413) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Bik was immunoprecipitated from 0.2 mg Ramos cell lysate with HA751413 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751413 at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: Ramos cell lysate (input) Lane 2: HA751413 IP in Ramos cell lysate Lane 3: Rabbit IgG instead of HA751413 in Ramos cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 20 seconds; ECL: K1801