Immunofluorescence staining CRISPR-Cas9 in CRISPR-Cas9 transfected 293T cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

Immunocytochemistry analysis of 293T cells transfected with or without px459 labeling CRISPR-Cas9 with Mouse anti-CRISPR-Cas9 antibody (EM50804) at 1/1,000 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-CRISPR-Cas9 antibody (EM50804) 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.