Hello,
To characterise the in vivo 32P-radiolabelling characteristics of
different cellular pools of lysophosphatidic acid, I'm trying to
develop a method for the separation of LPA species by reversed-phase
TLC. J.C. Darnell et al (Biochim. Biophys. Acta (1991), 1084:
269-299)
have done a separation of lyso-PI species on C-8 RP-TLC plates using
2
developments in 50% ethanol. This resulted in the separation of 16:0,
18:0 18:1 and 18:2 containing LPI's (and even of SN1- and SN-2
lyso's). The reproduceability is not entirely convincing though. The
main species of my LPA to be separated contain 16:0, 18:1 and 18:3
acyls.
Does anyone have experience/ suggestions on this? Would it be
necessary to neutralize the phosphate groups by including
counterions?
Should I try HP-RPTLC plates? Or am I wasting my time with this
experimental approach.
Thanks,
Steven.