Immunocytochemistry analysis of HeLa cells labeling Strep-Tag II with Mouse anti-Strep-Tag II antibody (HA600038) at 1/1,000 dilution. HeLa cells, transfected with Strep-Tag II-tagged Histone H3.1 (C-terminal) or ACAT2 (N-terminal) expression vector, respectively, were fixed in 4% paraformaldehyde for 10 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 Mouse anti-Strep-Tag II antibody (HA600038) at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) 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, green) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) were used as the secondary antibody at 1/1,000 dilution.

Flow cytometric analysis of HeLa cells labeling Strep-Tag II. HeLa cells, transfected with Strep-Tag II-tagged empty control or ACAT2 (N-terminal) expression vector, respectively, were fixed and permeabilized. Then stained with the primary antibody (HA600038, 1μg/mL) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).