Western blot assay was performed as previously described. 24 Briefly, the total protein of glioma tissues was extracted using a protein extraction kit (Beyotime). Equivalent amounts of protein (25‐50 μg) were then electrophoresed and transferred to polyvinylidene fluoride membranes (0.45 μm; Millipore). After being blocked, membranes were incubated with antibodies against ATF6 (24169‐1‐AP, Proteintech), EIF2α (sc‐133132, Santa), p‐EIF2α (# 3398S, CST), p‐IRE1α (sc‐390960, Santa) and GAPDH (60004‐1‐Ig, Proteintech) at 4°C for 16 hours. The membranes were then incubated with appropriate secondary antibodies (ProteinTech). Protein bands of interest were detected and quantified using a chemiluminescence kit (Beyotime), the ChemiDoc™ Touch detection system (Bio‐Rad Laboratories) and Image J software (National Institutes of Health).

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.