Cells fixed with 4% paraformaldehyde in PBS, fixed mouse brain samples, or postmortem human temporal lobe sections were blocked with a solution containing 5% normal donkey serum (Jackson ImmunoResearch Laboratories, West Grove, PA, USA), 2% BSA (Sigma-Aldrich), and 0.1% Triton X-100 (Sigma-Aldrich) for 1 hour at room temperature. Samples were then incubated with combinations of primary antibodies against RNF146, pAkt1, TH, or PAR, depending on the experiment, at 4°C overnight. Then, the fixed samples were washed with PBS containing 0.1% Triton X-100 and incubated with corresponding secondary antibodies conjugated with fluorescent dye at room temperature for 1 hour. Fluorescent images were obtained using a fluorescence microscope (Axiovert 200M, Zeiss, Oberkochen, Germany) or confocal microscope (LSM 710, Zeiss).

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.