distilled water shock of red blood cells - (Aug/25/2001 )
I am looking for a protocol for getting rid of red blood cell from tissue homogenates ( by enzyamatic digestion) .I am informed that to do FACS analysis with that tissue the red blood cells have to be got rid of and can be done so bydistilled water shock therapy of red blood cells.Can anyone provide me with a workable protocol.Thanks in advance.sankar
You can try to use the ammonium chloride to do so. Just like to follow the protocol to lyse the RBC from the PBMC
Are the cells adherent? If so you can add dH2O for about 1 minute and watch the RBC lyse (from phase bright & crenated to dark and ruptured) using an inverted microscope. Then you can just wash of the lysed cells and collect the cells of interest with a cell scraper.
The protocol I know is to incubate with distilled water for 10 seconds and then to quickly add 0.1 x volumes of 10 x PBS.
Worked fine in my hands.
You can lyse the red cell using lysis buffer
dissolve in water
adjust pH to 7.35 with 0.1NaOH
Total volume 500ml
filter it and kept at room temp.
Add the lysis buffer to the tissue homogenates and incubate for 5min at room temp.
Then neutralize the lysis buffer with PBS/EDTA.
I hope this may help.