I am looking to lyse some mammilian cells (A549) in order to quantify the amount of an anti-cancer drug (paclitaxel) by HPLC. Since my mobile phase consists in part of methanol, and paclitaxel which is highly insolubilbe in water but is solubile in methanol I would like to use methanol to lyse the cells so that I won't have problems preparing standard solutions and/or using an internal standard. Will simply methanol alone, given 10-15 minutes at room temperature be enough to lyse the cells? I have seen some talk of "acidified methanol" for bacterial cell lysis but nothing on the use of methanol alone for human cells.
Thanks for the help!
Using Methanol For Cell Lysis
1 reply to this topic
Posted 13 August 2011 - 08:05 PM
Methanol alone will fix mammalian cells - people use it as a fixative for immunofluorescence. It does this by precipitating the proteins. It may be enough to dissolve the cell membrane though, and this should release any dissolved paclitaxel (assuming it isn't complexed with proteins).