The cells were lysed with 50[micro]M digitonin
followed by addition of 1mM Ca[Cl.
Three-drug combinations of doxorubicin with lignans plus digitonin
Permeabiliszation and cell viability of PFFs treated with digitonin
were assessed by 0.
3-[(3-Cholamidopropyl)-dimethylammonio]-1-propanesulphonate (CHAPS) and 3-[(3-cholamidopropyl)-dimethylammonio]-2-hydroxy-1-propanesulphonate (CHAPSO) were from Calbiochem (Darmstadt, Germany); N-(2-hydroxyethyl)-piperazine-N'-(2-ethanesulphonic acid) (Hepes), tris(hydroxymethyl)aminomethane (Tris), ethylene glycol-bis([beta]-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), 1,6-diphenyl-1,3,5-hexatriene (DPH), sodium dodecyl sulphate (SDS), phenylmethylsulphonyl fluoride (PMSF), aprotinin, benzamidine, digitonin
, cholic acid, deoxycholic acid, n-dodecyl-[beta]-D-maltoside (DBM), and dodecylpoly[(ethylene-glycolether).
2+] to totally quench the Fura-2 fluorescence after the addition of digitonin
(50 taM), according to the method described by Sage (19).
14]C]-radio-labeled VLCFA substrates and one of two available methods: either in intact human skin fibroblasts cultured in monolayer (5); or in isolated fibroblasts permeabilized with digitonin
The first colorimetric assay for cholesterol in serum, reported by Grigaut in 1910, combined purification by digitonin
with an acid reagent.
Combination of the saponin digitonin
(5 [micro]M), which also inhibits P-gp, with SM significantly enhanced the inhibition of P-gp activity.
The comparative method was based on the use of 1 g/L digitonin
for 35 s before centrifugation and resuspension of the pellet in MOPS buffer (12).
Briefly, membrane and cytosolic fractions were preincubated with the salt solution for 10 min at 37 [degrees]C and incubated for another 15 min in the presence or absence of 100 [micro]M PKC-specific peptide substrate, subsequently with 50 [micro]g/m1 digitonin
and 100 [micro]M ATP mixed with [[gamma]-[.
in phosphate-buffered saline (PBS), at final concentrations of 0, 20, 45, and 90 [micro]mol/L, was used to control the degree of whole-blood hemolysis (21).
Treatment with 40[micro]M digitonin
increased the number of parasites marked with PI by 68.