Equal amounts of extracted proteins (20 to 40 μg) were resolved on 10% SDS–polyacrylamide gel electrophoresis gels, and the proteins were transferred to nitrocellulose membranes (Whatman). The membranes were incubated with TGFBR1 (1:1000, 3712; Cell Signaling Technology), SMAD2 (1:1000, 3102; Cell Signaling Technology), pSMAD2 Ser465/467 (1:1000, 138D4; Cell Signaling Technology), SMAD4 (1:1000, 46535; Cell Signaling Technology), and β-actin (1:5000, 3700; Cell Signaling Technology) or β-tubulin (1:5000, 2146; Cell Signaling Technology) followed by incubation with appropriate horseradish peroxidase–conjugated antibodies at room temperature for 1 hour. Proteins were detected by an enhanced chemiluminescence solution (Thermo Fisher Scientific).

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.