The proteins were analyzed from whole-cell lysates of BMSCs cultured with OM after 1 week. BCA protein assay reagent (Boster, AR0197, Wuhan, China) was used to measure the protein concentration. Next, 40-μg protein samples were separated by SDS-polyacrylamide gels and transferred to PVDF membranes. The membranes were then blocked with 5% bone serum albumin (BSA) for 1.5 h and incubated with primary antibodies (COL1, BMP2 at 1:1000 dilution, β-actin at 1:5000 dilution, Abcam, UK; OCN at 1:1000 dilution, Santa Cruz Biotechnology) at 4 °C overnight. Blots were then incubated with secondary antibodies for 1 h before they were detected by the Western ECL Substrate Kit (Thermo Pierce, USA), while the proteins were normalized by β-actin.

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.