Blocking buffer: 5% NFDM/TBST Primary ab dilution: 1:10000 Primary ab incubation condition: 2 hours at room temperature Secondary ab: Goat Anti-Mouse IgG H&L (HRP) Lysate: 1: HeLa, 2: SH-SY5Y, 3: C2C12 Protein loading quantity: 20 μg Exposure time: 3 s Predicted MW: 43 kDa Observed MW: 43 kDa

Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (WNT10B) Monoclonal Antibody, Unconjugated (bsm-60528M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.

Tissue: Human liver cancer 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: 1:100 Primary ab incubation condition: 1 hour at room temperature Secondary ab: SP Kit(Mouse)(sp-0024) Counter stain: Hematoxylin (Blue) Comment: Color brown is the positive signal for bsm-60528M

Cell line: HeLa Fixative: 4% Paraformaldehyde 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-60528M