Sigma-Aldrich
Red Blood Cell Lysing Buffer Hybri-Max
R7757
We use this pre-made red blood cell lysis buffer during our mouse and human preps for flow cytometry. Having a pre-made solution saves us a lot of time and getting rid of these cells is really important for our flow cytometry protocol-less unnecessary cells means less time sorting, and less time sorting is always great!
Flow cytometry prep
Mouse or human single cell suspension
For mouse lung cells-we use this right after our digest at about 1ml RBC lysis buffer per 10 million cells (we eyeball total cell number, so this is just an estimate)For human-we use this after we've gotten our tissue down to a single cell suspension and use about the same concentration as mouse, probably a bit more because we generally have more red blood cells in our human preps. Sometimes we will do a second lysis step right before staining as well, depending on what our cell pellets look like.We usually lysis for 3-5 minutes on a rocker at room temperature. This stuff is strong, so definitely don't forget about it! I also like to lyse in a tube that has enough space for me to dilute the buffer before spinning down. I like to get the cells out of the lysis as fast as possible because it's harsh on them.
Filter single cell prep after lysing to keep it nice and clean!
Really clean pellets free of red blood cells. This helps us keep our flow preps clean and efficient and saves us a lot of time sorting and making rbc lysis buffer.
N/A
Works well consistently
None
Helpful step in our flow cytometry prep that saves us time and is super easy to do!