Blocking buffer: 5% NFDM/TBST Primary Ab dilution: 1:1000 Primary Ab incubation condition: room temperature 2h Secondary Ab: Goat Anti-Mouse IgG H&L (HRP) Lysate: 1: MCF-7, 2: T47D, 3: MDA-MB-231 Protein loading quantity: 20 μg Exposure time: 60 s Predicted MW: 15 kDa Observed MW: 15 kDa

Cell line: HepG2 Fixative: 4% Paraformaldehydel Permeabilization: 0.1% TritonX-100 Primary Ab dilution: 1:50 Primary incubation condition: 4°C overnight Secondary Ab: Goat Anti-Mouse IgG Nuclear counter stain: DAPI (Blue) Comment: Color green is the positive signal for bsm-60551M

HepG2 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (FIS1) monoclonal Antibody, Unconjugated (bsm-60551M) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Mouse IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.