Camptothecin (20 [micro]M) and digitonin
(80 [micro]M) were used as positive controls for apoptosis and necrosis, respectively (PC), and cells not treated with apigenin were used as negative controls (NC).
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
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
(5) Cells were washed with PBS and permeabilized with 0.025% digitonin
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.
The differential detergent fractionation method described by Rambsy and Makowski in 1999, was modified using a digitonin
(0.015%) buffer to permeate HTR8/SVneo human trophoblast cells and obtain the cytoplasm proteins in the soluble fraction, while membrane proteins were dissolved from the pellet with Triton X-100 (0.5%) buffer.
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.