The molecular weights of the respiratory complexes or supercomplexes were estimated by using digitonin
bovine heart mitochondrial complexes as standard: single complex: I = 1,000 kDa, V = 750 kDa, III2 = 500 kDa, IV = 230 kDa, II = 130 kDa; supercomplexes: I-III-[IV.sub.1-4] = 1500-2100 kDa, [V.sub.2] = 1500 kDa.
The cells were then pelleted by centrifugation for 3min at 1700g and 4[degrees]C; gently suspended in permeabilization buffer (220 mM mannitol, 68 mM sucrose, 80 mM KCl, 10 mM HEPES (pH7.4), 2.5 mM EGTA, 1 mM EDTA, 1 mM DTT, 200 (g/ml digitonin
, and 1% protease inhibitor cocktail); placed on ice for 5 min; and centrifuged for 5min at 750g and 4[degrees]C.
Osteosarcoma cybrids were permeabilized with digitonin
(4 [micro]M), and respiration was initiated with complex I substrate (malate/pyruvate, 5 mM).
(80 [micro]M) was used as a positive control, and untreated cells were used as negative controls.
The positive control was cells that were incubated with 50 [micro]g/mL digitonin
(Amend Drug and Chemicals Co., USA) for 5 min, as previously described (30).
After treatment, the cell pellet after centrifugation was lysed with Tris buffer (50 mol/L Tris-HCl, 1 mmol/L EDTA, and 10 mmol/L EGTA at pH = 7.4) containing 10 [micro]mol/L digitonin
for 20 min at 37[degrees]C.
Synergism of three-drug combinations of sanguinarine and other plant secondary metabolites with digitonin
and doxorubicin in multi-drug resistant cancer cells.
Frischknecht, "Synergistic and additive effects of epigallocatechin gallate and digitonin
on Plasmodium sporozoite survival and motility," PLoS ONE, vol.
Test for digitonin
sensitivy and sterol requirement.
Behavior of polymer supported digitonin
with cholesterol in the absence and presence of butter oil.
However, its cells are sensitive to digitonin
. The presence of only plasma membrane makes it vulnerable to osmotic shock and the effect of detergents [21, 29].
sensitivity test was carried out to differentiate between Mycoplasma and Acholeplasma genera using filter paper discs impregnated with 0.2 mL of 1.5% (W/V) ethanol solution of digitonin
and dried overnight.