Blocking buffer: 5% NFDM/TBST Primary ab dilution: 1:1000 Primary ab incubation condition: 2 hours at room temperature Secondary ab: Goat Anti-Rabbit IgG H&L (HRP) Lysate: 1: A431, 2: Mouse skin, 3: Rat skin Protein loading quantity: 20 μg Exposure time: 10 s Predicted MW: 45 kDa Observed MW: 45 kDa

Tissue: Human prostatic hyperplasia Section type: Formalin fixed & Paraffin -embedded section Retrieval method: High temperature and high pressure Retrieval buffer: Tris/EDTA buffer, pH 9.0 Primary ab dilution: 1:1000 Primary ab incubation condition: 1 hour at room temperature Secondary ab: SP Kit(Rabbit) (sp-0023) Counter stain: Hematoxylin (Blue) Comment: Color brown is the positive signal for bsm-60577R

Tissue: Human esophagus Section type: Formalin fixed & Paraffin -embedded section Retrieval method: High temperature and high pressure Retrieval buffer: Tris/EDTA buffer, pH 9.0 Primary ab dilution: 1:1000 Primary ab incubation condition: 1 hour at room temperature Secondary ab: SP Kit(Rabbit) (sp-0023) Counter stain: Hematoxylin (Blue) Comment: Color brown is the positive signal for bsm-60577R

Cell line: A431 Fixative: 100% Ice-cold methanol Permeabilization: 0.1% TritonX-100 Primary ab dilution: 1:50 Primary incubation condition: 4°C overnight Secondary ab: Goat Anti-Rabbit IgG Nuclear counter stain: DAPI (Blue) Comment: Color green is the positive signal for bsm-60577R

Cell line: A431 Fixative: 4% Paraformaldehyde Permeabilization: 0.1% TritonX-100 Primary ab dilution: 1:100 Secondary ab: Goat anti Rabbit IgG Unlabelled control: The cell without incubation with primary antibody and secondary antibody (Black line). Isotype control: Rabbit monoclonal IgG (Blue line). Comment: Line red is the positive signal for PTM-5986