6]) were washed with ice-cold Dulbecco's phosphate-buffered saline and were resuspended in 100 [micro]L of ice-cold digitonin
lysis buffer (0.
Behavior of polymer supported digitonin
with cholesterol in the absence and presence of butter oil.
These vacuoles were positive for Sudan III, Sudan black B, oil red O, Nile blue, periodic acid-Schiff, Schultz test, and digitonin
stain, and were negative for performaric acid--Schiff stains.
6 [micro]mol/L digitonin
for the permeabilization of the cell membranes, 1 mmol/L pyruvate, 1 mmol/L malate, and, where indicated, 2.
Fura-2AM, serotonin, DNA, DNaseI, guanidine hydrochloride, digitonin
and EGTA were purchased from Sigma-Aldrich Chemicals, Bangalore, India.
Germinally transgenic tadpoles were produced by restriction enzyme-mediated integration nuclear transplantation according to Kroll and Amaya (1996), with the following modifications: sperm was purified by centrifugation on a two-layer discontinuous Percoll (Sigma) gradient before the permeabilization step, which was performed with digitonin
(Sigma) instead of lysolecithin.
Kinetics of the inactivation of the muscarine cholinoreceptor, solubilized by digitonin
Additionally, in threedrug combinations of lignans with the saponin digitonin
and doxorubicin MDR reversal activity was even stronger enhanced.
Permeabilization and cell viability of PFFs treated with digitonin
The muscarinic acetylcholine receptors (mAChR), which are members of the family of receptors that are coupled to guanine nucleotide binding proteins (G proteins) and whose structure is characterized by the seven membrane-spanning [alpha]-helical domains [2, 3], are usually solubilized in digitonin
or in a mixture of digitonin
and sodium cholate, which have yielded more than 50% of the soluble receptors with retention of ligand binding activity [4, 5].
The results were expressed as the percentage of ATP released relative to the total ATP present in cells lysed by means of digitonin
(50 [micro]M) (16).
min] values were determined by addition of digitonin
(50 [micro]M) and [Mn.