Immunocytochemistry analysis of MDA-MB-231 cells labeling Kindlin 2 with Mouse anti-Kindlin 2 antibody (HA601343) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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 Mouse anti-Kindlin 2 antibody (HA601343) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

Kindlin 2 was immunoprecipitated from 0.2 mg MDA-MB-231 cell lysate with HA601343 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA601343 at 1/2,000 dilution. Anti-Mouse IgG for IP Nano-secondary antibody (NBI02H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: MDA-MB-231 cell lysate (input) Lane 2: HA601343 IP in MDA-MB-231 cell lysate Lane 3: Mouse IgG instead of HA601343 in MDA-MB-231 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 38 seconds; ECL: K1801