Immunocytochemistry analysis of U-937 cells labeling PI 3 Kinase p85 alpha with Mouse anti-PI 3 Kinase p85 alpha antibody (HA610084) 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-PI 3 Kinase p85 alpha antibody (HA610084) 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.

Western blot analysis of PI 3 Kinase p85 alpha on different lysates with Mouse anti-PI 3 Kinase p85 alpha antibody (HA610084) at 1/5,000 dilution. Lane 1: A549-si NT cell lysate Lane 2: A549-si PI 3 Kinase p85 alpha cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 84 kDa Observed band size: 84 kDa Exposure time: 2 minutes; ECL: K1802; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA610084) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.