Approximately 1 × 106 cells were stained with 5–10 μg/ml of CD45-Pacific Blue or 2.5 μl of CD71 stock solution for 30 min at 4–8 °C in the dark. Cells were washed twice with PBS by centrifugation at 1455×g for 4 min, at 4 °C. After the last wash 200 μl of FACS buffer was added to the cells.

Double staining with SYBR Green I and CD45-Pacific Blue: A volume of 30 μl of SYBR Green I at 10× was added to pelleted cells that were previously stained with CD45-Pacific Blue (as mentioned above) for 30 min at 4–8 °C, in the dark. After incubation a volume of 200 μl of FACS buffer was added.

Triple staining with SYBR Green I, CD45-Pacific Blue and CD71-APC: Approximately 1 × 106 cells were stained with 10 μg/ml of CD45-Pacific Blue, 2.5 μl of CD71 stock solution and 30 μl of SYBR Green I at 10x. Samples were incubated for 30 min in the fridge (4–8 °C) in the dark. Cells were washed twice with PBS by centrifugation at 1455×g for 4 min, at 4ºC. After incubation a volume of 200 μl of FACS buffer was added.

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.