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Blocking buffer: 5% NFDM/TBST Primary ab dilution: 1:2000 Primary ab incubation condition: 2 hours at room temperature Secondary ab: Goat Anti-Rabbit IgG H&L (HRP) Lysate: 1: C2C12 cell lysate, the Ly6A Rabbit mAb pre-adsorbed with 3μM of the synthetic peptides; 2: C2C12 cell lysate, the Ly6A Rabbit mAb with no peptide blocking. Protein loading quantity: 20 μg Exposure time: 10 s Predicted MW: 14 kDa Observed MW: 13 kDa

Blocking buffer: 5% NFDM/TBST Primary ab dilution: 1:2000 Primary ab incubation condition: 2 hours at room temperature Secondary ab: Goat Anti-Rabbit IgG H&L (HRP) Lysate: 1: Mouse thymus, 2: C2C12 Protein loading quantity: 20 μg Exposure time: 60 s Predicted MW: 14 kDa Observed MW: 13 kDa

Tissue: Mouse testis 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(Rabbit) (sp-0023) Counter stain: Hematoxylin (Blue) Comment: Color brown is the positive signal for bsm-60617R