Anti lyz1 antibody

Total protein from tissues was using RIPA Buffer (Thermo Fisher Scienti c, Inc.) supplemented with protease inhibitor cocktail (Roche Applied Science). Protein samples (40 µg/sample) for each group was loaded and resolved on a 10% SDS-PAGE gels, and subsequently transferred to the polyvinylidene uoride (PVDF) membranes membrane (EMD Millipore). Then, the membranes were blocked with 5% skim milk at room temperature for 1 h and incubated at 4˚C overnight with primary antibodies: anti-SI antibody (1:400; cat no. ab84977), anti-Tff3 antibody (1:400; C cat no. sc398651), anti-Lyz1 antibody (1:250; cat no. ab189937), anti-ChgA antibody (1:200; cat no. ab15160), anti-Rspo1 antibody (1:1,000; cat no. ab106556), anti-Rspo2 antibody (1:1,000; cat no. ab132836), anti-Rspo3 antibody (1:1,000; cat no. ab233113), anti-Rspo4 antibody (1:1,000; cat no. ab189515), and anti-β-actin antibody (1:1,000; cat no. ab8226) (all from Abcam, Inc.). The blots were incubated with the horseradish peroxidase-conjugated secondary antibodies (cat no. 7074S; Cell Signaling Technology, Inc.) at 37˚C for 1 h at room temperature and visualized using an enhanced chemiluminescence Ultra Western HRP Substrate kit (cat no.WBULS0100; EMD Millipore). Signals were analyzed by Quantity One software version 4.6.2 (Bio-Rad Laboratories, Inc.) and the intensity values were normalized to β-actin.