Western blotting was performed using the Invitrogen Novex Mini-Cell system with NuPAGE 4 to 12% bis-tris gels. Proteins were transferred to nitrocellulose membranes (0.45 μm; Bio-Rad) and blocked using LI-COR blocking buffer at a 1:1 ratio with PBS for 1 hour at room temperature. Primary antibodies were mixed in tris-buffered saline (TBS)–0.1% Tween 20 (TBST) with 2% BSA and incubated with membranes overnight at 4°C on a shaker. Membranes were then washed three times for 10 min in TBST and incubated with fluorophore-conjugated secondary antibodies in TBST with 2% BSA for 1 hour at room temperature. Membranes were washed three times for 10 min in TBST and imaged using a LI-COR Odyssey imaging system. Quantification of bands was done using ImageStudio software (LI-COR).

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.