Sds page gel preparation kit

Vital organs removed from normal mice were crushed with a mortar using liquid nitrogen under ice cold conditions and lysed with RIPA lysis buffer (YEASEN, #20101ES60) and InStab^™ Protease Cocktail, EDTA-free, mini, tablet-form (YEASEN, #20123ES10). After centrifugation at 14,000 r/min at 4 °C for 20 min, supernatants were collected and protein concentration was determined using the Pierce^™ BCA protein assay. All samples were heated for 10 min at 95 °C in 5× SDS-PAGE Protein Loading Buffer (YEASEN, #20315ES05) before loading. Cell lysate aliquots were separated in 10% SDS-PAGE (SDS-PAGE Gel Preparation Kit, YEASEN, #20328ES50). Proteins were electro-blotted onto polyvinylidene fluoride membranes (Millipore) and blocked by incubation with TBST (TBS and 0.2% Tween-20) containing 5% non-fat dry milk for 1 h at room temperature. After blocking, membranes were washed in TBST three times for 5 min and then probed with the Recombinant Anti-ACE2 Momoclonal Antibody (Abcam, #ab108252) overnight at 4 °C. Subsequently, the membranes were washed with TBST and incubated for 1 h at room temperature with goat-anti rabbit-HRP (Servicebio, #GB23303) in TBST. Finally, membranes were washed three times and enhanced with Clarity^™ Western ECL substrate (YEASEN, #36208ES60), followed by imaging with Alliance Micro Q9 (UVITEC).