The ultrastructural localization of LC3B was examined using zebrafish larvae, employing the post-embedding method as described previously49,50. Small larvae specimens embedded in LR White Resin, prepared as semi-thin sections, were used. The RFP antibody was used for the detection of mCherry protein, because it reacts with RFP and other RFP variants, such as mCherry. Ultra-thin sections (70 nm thick) were cut, incubated with a rabbit polyclonal LC3B antibody (1:300) and a mouse monoclonal RFP antibody (1:100) for 2 h at 24 °C , and reacted with 10-nm gold colloidal particle-conjugated anti-rabbit IgG (EMGFAR10; British BioCell International, Cardiff, UK; 1:30) and 5-nm gold colloidal particle-conjugated anti-mouse IgG (EMGMHL5; British BioCell International; 1:30). Finally, the sections were stained with lead citrate and examined using a JEM-1400 electron microscope at 80 kV (JEOL, Tokyo, Japan).

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

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.