THE UNIVERSITY OF MANITOBA STUDIES ON OVINE PLACINTAL ...

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(c) À.nalvticaL .{rod.iu¡n dodêcyl Eulfatêj polyacrl.taniiiê gê1 (slab)
ê1ec troÞhorê s Ls
Polyacrylanide. gel eLectrophoresis contaÍning
sodium dodecyl sulfate was carried out as described by
t{eber and Osborn (1969) with slight modification. For the
resolving gel , a 129 acrylamide was used, whereas for the
stacking gel-, a 3t acrylamÍde was used. Elect,rophore sis
was al"lowed to proceed at a constant current. of 20 mitli-
amperes. The samples were boiled at 90 c for 'l minute with 2% sodium
dodecyi sulfate, 2% glycerol , and sma.ll amount of bromophenol bìue before
bein,g, applíed to the top of the stacking gel . For eaah,sanp,¡e, l0 ug of protein
was used.' .
The staining was perforned by placing the. gel in
508 TCA containing 0.25t Coornassie Brillant Blue R250 for
2-3 hours. Then the stained 9e1 v¡as removed from the staining
solution and destained in destaíning solution containing
l0 g nethanol and 5* acetic ì.acid.
(d) MolecuLar wêight. detêrminat.ion:
The molecular weight of opL was estimated by a
cornpar is ion of the rnobility of purified opï, wiÈh the mobility
of p.roteins of known mofecular welight.in 12? polyâctyla¡nide
gels containing sodium dodecyl sulfate as described above.