Triton solution preparation

Much of the adenylate cyclase activity of mammalian tissues can be solubilised or dispersed in Triton solution [18]. Particulate preparations can be stored at -70°C for long periods without loss of activity, but at higher temperatures the activity from cells of multicellular organisms is rapidly lost [6]. 

Experiments Designed to Determine the Effect of Water Additives on Count Rates. The composition of the scintillator solution was 70 Vol % toluene and 30 Vol % Triton to which 7g/l PPO and 0.35g/l DMPOPOP were added. To each vial containing 10 ml of this solution was added the desired amount of water followed by the addition of 10 yl of the radioactive sample, H-toluene, H-H20, C-cyclohexane, C-sodlum carbonate and NaCl, the two latter compounds dissolved in a -microamount of H2O. The counting solutions prepared in this way were vigorously shaken and counted at room temperature. 

Triton X-100, like gelatin, suppresses both positive and negative maxima, but, unlike gelatin, its aqueous solution is stable. A stock 0.2 per cent solution is prepared by shaking 0.20 g of Triton X-100 thoroughly with 100 mL of water. About 0.1 mL of this solution should be added to each 10 mL of the sample solution to give a Triton X-100 concentration of 0.002 per cent. 

Epoxyeyclohexanone has been prepared in 30% yield4 by epoxi-dation of 2-cyclohexen-l-one with alkaline hydrogen peroxide, using a procedure described for isophorone oxide (4,4,6-trimethyl-7-oxabicyclo[4.1.0]heptan-2-one).5 A better yield (66%) was obtained using f r/-butyl hydroperoxide (1,1-dimethylethylhydroperoxide) and Triton B in benzene solution.6 The procedure described here is simple and rapid. 

After 2 h incubation of the prepared antibody beads with UV-crosslinked extract in a cold room, the beads are washed 4 x with 100 /A RIPA buffer (50 mMTris-HCl pH 7.5, 150 rnMNaCl, 1% NP-40, 0.5% sodium deoxycholate, and 0.1% SDS) and lx with genomic DNA lysis buffer (50 mM Tris, pH 7.4, 10 mM EDTA, 500 mM NaCl, 2.5 mM DTT, 0.5 mM spermidine, 1% Triton X-100). Approximately 300 /(I of PK solution (1 mg/ml proteinase K in genomic DNA lysis buffer and 0.2 U//A RNase inhibitor) is added to the total lysate previously kept on ice and the beads are then incubated at 37° for 30 min. Gently flick the tubes to resuspend the beads every 10 min during the incubation. After removal of the proteinase K solution, 300 /A of RNA extraction solution (4 M guanidine thiocyanate, 0.5% sarkosyl, and 25 mM sodium citrate, pH7) is added to the beads, incubated for 10 min and the supernatant is mixed with 30 fig yeast tRNA (as a carrier) and 30 fil of 3 M sodium acetate. The RNA solution is phenol-chloroform extracted, ethanol-precipitated, and the pellet washed once with 70% ethanol. The dry pellet is used for 1st strand cDNA synthesis, followed by PCR analysis. The removal of proteins... 

Prepare a solution consisting of 2.5 ml glycerol, 100 pi of 10 percent Triton X-100 (Thermo Fisher Surfact-Amps X-100), and 10 pi 2-mercaptoethanol. 

A Working Solution of the stain should be prepared just before use, by combining 10 ml of 0.1% Triton X-100 in phosphate-buffered saline, 2 mg RNase (DNase-free, Sigma), and 200 pi PI solution (Sigma, 1 mg/ml in distilled water). Stock solutions of Triton X-100 and PI should be stored at 4°C. Centrifuge the ethanol-fixed cells (200 x g for 5 min) and discard the supernatant, removing it completely. 

In order to more closely represent the volatilization environment that would be encountered in an evaporation pond, Triton X-100, a non-ionic emulsifier similar to those used in some pesticide formulations, was added to prepared pesticide solutions at 1000 ppm. The presence of this emulsifier caused a decrease in the percent pesticide volatilized in one day in all cases except for mevinphos (Table VI). Three mechanisms are probably in operation here. First, Triton X-100 micelles will exist in solution because its concentration of 1000 ppm is well above its critical micelle concentration of 194 ppm (30). Pesticide may partition into these micelles, reducing the free concentration in water available for volatilization, which will in turn reduce the Henry s law constant for the chemical (31). Second, the pesticides may exhibit an affinity for the thin film of Triton that exists on the water surface. One can no longer assume that equilibrium exists across the air-water interface, and a Triton X-100 surface film resistance... 

Triton X-100, EDTA, dithiothreitol and electrolyte protect enzyme in dilute solution and against denaturation by heat or extreme pH-values [12, 48] <2>, at low dithioerythritol concentrations enzyme tends to aggregate [5] <2>, bovine serum albumin, 1 mg/ml, stabilizes dilute enzyme solutions [5] <2>, diadenosine pentaphosphate, i.e. AP5A, stabilizes during preparative electrophoresis [7]... 

Functional mesostructures were prepared by simultaneously adding both TEOS and MPTMS to solutions of various surfactants, including alkylamines (octylamine and dodecylamine) and non-ionic surfactants (Tergitol 15-S-12 and Triton-X). The exact conditions for these synthesis procedures have been described in other publications.19,21 The mixtures were stirred for 24 hours and the products filtered, air dried and washed free of the assembly surfactant by Soxhlet extraction over ethanol for 24 hours. 

As for Basic Protocol 1, the LOX enzyme is prepared according to the Support Protocol. However, more care should be taken to reduce turbidity and other UV-absorbing materials caused by suspended lipid and pigments, especially when low activity is expected (requiring more enzyme addition). Triton X-100 is a UV absorber, and it causes more lipid and pigments to be suspended. Partial purification may be necessary. HEPES and PIPES buffers are not used in this method because of excessive absorption at 234 nm (0.5 to 0.6 absorbance for 0.1 M solutions) MES is useful with limitations (0.2 absorbance for 0.1 M solution). 

Precautions should also be taken with the aqueous solvents used to prepare standard working solutions in order to avoid adsorption problems, especially at low concentrations. To minimize adsorption during standard curve and validation sample (VS)/QC preparations, aliquots of the high-concentration stock solutions should be spiked promptly into the control blank plasma to prepare the standards and VS/QC. Once the compounds are in an environment of protein solutions, the adsorption problem is negligible. If there is a problem, however, adding or pre-rinsing with a solution of protein or a chaotropic agent (e. g., Tween, Triton X-100 or CHAPS) may help to alleviate the problem. 

The preparation of milk samples as slurries to be analyzed by ICP-AES was addressed by Carrion et al. [33]. The samples were emulsified with w-ethoxy nonylphenol and then diluted 10-fold with 1 percent HNO3. Aqueous solutions with the same amount of emulsifier and acid were used as calibration standards. In another work samples of milk and formulae were prepared as slurries and were then analyzed using aqueous standards [7], Dean et al. [34] used ICP-MS to determine Pb isotope ratios in milk powders just suspended in a diluted solution of Triton X-100. 

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