THE UNIVERSITY OF MANITOBA STUDIES ON OVINE PLACINTAL ...

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for 30 min. Then the precipitate ¡rqs ¡esuspe¡ded in 2 rnl of 5%
TCA, and ¡e.cent¡ifuged. Finally, the precipitate was dissolved in
0.5 ml of 0.5N Na0H, re-precipitated with 5/ TCA, recentrifuged,
and the precipÍtate was dissolved in O.l t'l Na0H and counted in
Aquasol II as described above.
(5) Chromatography of the incubation medium
3 ml of incubation medium was chromatographed on a
Sephadex G-100 column (45 X Z cm) whìch was equilibrated and
eluted with 0.01 M pBS conta.ining SmM .leuclne. Fractions of 3 ml
were collected. The effluent fractions were assayed for total
oPL by radioimmunoassay, 3H-oeL by inrmunopreci pi tatì on , 3H_protein
by TCA precipitation, and total protein concentrafion by measuring
the absorbance at 278 nm.