The negative control group (untreated cells) contained viable cells (Figure 2A-C), while the group treated with
digitonin exhibited dead cells (Figure 2D-F).
isolates showed inhibition zones > 3 mm in the
digitonin sensitivity test.
Briefly, a known amount of tissue homogenate is added to 0.01% chilled
digitonin and was centrifuged at 4[degrees]C and 0.05 mL of the supernatant of the tissue homogenate is mixed with 3 mL of [H.sub.2][O.sub.2] phosphate buffer.
An aliquot of each Folch extract was evaporated and the residue dissolved in 50 mM sodium phosphate buffer (pH 7.4) containing
digitonin 1%.
(5) Cells were washed with PBS and permeabilized with 0.025%
digitonin (Sigma).
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.
1 - - ++ - ++ ++ NE NE NE Phenax - - ++ rugosus - - ++ NE NE NE Standards Kaempferol Gallic
Digitonin +++ Acid +++ +++ Family Species Phytocompounds Lactones Apocynaceae Mandevilla - veraguensis - - Blepharodon - mucronatum - - Mandevilla - fendleri - - Cactaceae Rhipsalis - micrantha - - Costaceae Costus sp.
The incubation volume in each well was 50 [micro]L, containing 5 [10.sup.4]L cell suspension (2-3 x [10.sup.5] cells), 100 nmol/L fluorescent probe, and a substrate mix consisting of 30 mmol/L potassium phosphate, 75 mmol/L potassium chloride, 8 mmol/L Tris, 1.6 mmol/L EDTA, 5 mmol/L [MgCl.sub.2], 0.2 mmol/L [p.sup.1],[p.sup.5]-di(adenosine-5') pentaphosphate (myo-adenylate kinase inhibitor), 32.6 [micro]mol/L
digitonin for the permeabilization of the cell membranes, 1 mmol/L pyruvate, 1 mmol/L malate, and, where indicated, 2.0 mmol/L ADP, pH 7.4 (1).
dodecyldimethylamine oxide, lysolecithins), and ii) detergents belonging to the group of bile salts or other steroid-based detergents such as 3-[(3-Cholamidopropyl) dimethylammonio]-1-propanesulfon ate (CHAPS) and
digitonin. The foremost challenge in the solubilization of a membrane protein is to obtain a soluble protein in a stable form that retains its original function sodium dodecyl sulfate (SDS) is a detergent that is very much used in the procedures of [Na.sup.+]/[K.sup.+]-ATPase solubilization, although it partially denatures the enzyme molecule.
Fura-2AM, serotonin, DNA, DNaseI, guanidine hydrochloride,
digitonin and EGTA were purchased from Sigma-Aldrich Chemicals, Bangalore, India.