Pigments were extracted from thylakoids with 85% acetone buffered with Na2CO3 or from intact cells with dimethylformamide.
Wet season Chl a filters, from the Betsiboka basin only, were extracted with 1 ml of 90% acetone buffered with saturated MgCO3, after which they were incubated overnight at 4°C.
Six leaves from each sample (about 150 to 200 mg) were chopped into 2 3 mm pieces and placed into Precellys tubes CK28R with 1 mL of ice cold TCA-acetone precipitation buffer [10% TCA in acetone] and homogenised using Precellys for 45 seconds at 6500 rpm.
Frozen samples were homogenized with a Tissue Tearor Homogenizer (Model 395, Dremel, Mexico) in pure acetone solution buffered with CaCO3.
To further improve the organic stability of the enzyme, Kartal et al. (2011) suggested the mixing of acetone with another buffer.
The universal column has been replaced with Metrosep C4 150/4.0 column with 3.0 mM nitric acid buffer: Acetone: 90:10 (%v/v), the peaks of 1-deoxynojirimycin, miglitol, and castanospermin were separated but not resolved properly.
After thawing, the slides were fixed for 10 min in ice-cold acetone, washed with buffer and blocked for 10 min at room temperature (RT) with biotin blocking system part one.
Briefly, cells were rinsed twice with 1 mL of Tyrode's balanced salt solution (TBSS, Sigma-Aldrich) and then fixed in citrate buffered acetone for 30 seconds at room temperature.
SBP also showed maximum activity at different concentrations of acetone using a phosphate buffer, pH 6.0 than with the other pH buffers.
Tissue sections of 10 μm thickness were treated with ice-cold acetone, rehydrated in Tris buffer solution (50 m Tris, 100 m NaCl, 0.001% Aprotinin, pH 7.4), and blocked with Tris buffer solution 20% fetal calf serum.
Samples were extracted with water at temperatures of 20, 60 and 90 °C, using mixtures of methanol/water (9 1, 1 1 and 1 9), Tris buffer and acetone and the extracts speciated.
