Proteins in total cell lysates or nuclear fractions were separated with 7–15% SDS-polyacrylamide gel electrophoresis (PAGE) (Choi et al. 2019) and then transferred onto a PVDF membrane. To reduce non-specific antibody reactions, the PVDF membrane was first blocked with BSA. After washing twice with Tris-buffered saline including Tween 20 (TBST), the membrane was incubated overnight with primary antibody in BSA. After three washing steps with TBST, the membrane was probed with a secondary antibody conjugated with horseradish peroxidase in BSA for 1 h. Immunoreactive bands were detected using an enhanced chemiluminescence kit (Pierce ECL Western blotting substrate, Thermo Scientific, Waltham, MA). Two different blots were obtained from two independent Western blotting analyses. Band intensity was measured and quantified using ImageJ (Bethesda, MD).

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



Q&A
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.