Triton temperature

The scale of (D) is two times that of (C). No added Triton. Temperature, 293K. 

A five-step synthesis of ethyl ester of eyelie hydrazonie aeid 314 used in the synthesis of natural produets has been deseribed [94JCA(CC)1867]. The eonden-sation of methoxybutenone with EtCOaCN (t-BuOK, THF, —78°C) is eompleted with the formation of ketoester 310 in 72% yield. The addition of methanol to the latter (Triton B, MeOH, room temperature, 88%) and the reduetion with NaBH4 (EtOH, —78°C) leads to the aleohol 312, yield 90%. The dianion of 312 (EDA, THE, -78°C) reaets with t-butylazodiearboxylate (t-BuOaC—N=N—COaBu-t) to form adduet 313, the treatment of whieh with trifluoroaeetie aeid affords the ester 314 in 55% yield [94JCA(CC)1867]. 

Detection and result The dried chromatogram was heated in the drying oven at 180 °C for 20 min. After cooling to room temperature it was dipped twice for 1 s into a solution of Triton X-100 — chloroform (l-t-4) which stabilized the fluorescence and increased its intensity by a factor of 2.5. Between the two dipping steps the chromatogram was air-dried in the dark for 30 min until the chloroform had completely evaporated. 

Figure 6.3 Conparlson of the separation of the octylphenol poly(ethylene glycol) ether, Triton X-16S on a packed column, left, and an open tubular column, right, using UV detection. For the packed column separation al0cmx2mmI.D. column packed with Nucleosil C g, d. 3 micrometers, temperature > 170 C, and mobile phase carbon dioxide (2 ml/min] and methanol (0.15 nl/rnin). pressure programmed from 130 to 375 bar in 12 min were used. For the open tubular column separation a 10 m x 50 micrometers I.O., SB-Biphenyl-30, temperature = 175°C, mobile phase carbon dioxide (0.175 ml/min) and 2-propanol (0.0265 ml/min) pressure programmed, 125 bar for 5 min, then ramped from 125 to 380 bar over 19.5 min, and held at 380 bar for 15 min. were used. (Reproduced with permission from ref. 57. Copyright Preston Publications, Inc.) ...

The next day (48 h posttransfection), the medium is replaced with fresh DMEM containing different concentrations of compound (nM—fiM concentration range). After 12 h of incubation, the cells are washed with PBS, 40 1 of luciferase lysis buffer [100 mM KxP04 (pH 7.8), 0.2% Triton X-100] is added to each well, and the plate is incubated for 15 min at room temperature with gentle rocking. The cell extract is transferred into Eppendorf tubes and kept on ice. 

A study of by Palmer-Toy et al.,12 summarized in Table 19.1, provides further empirical evidence of the utility of techniques coupling heating with efficient protein extraction for the proteomic analysis of FFPE tissue. A specimen from a patient with chronic stenosing external otitis was divided in half and preserved as fresh-frozen tissue or FFPE. Ten micromolar sections of the FFPE tissue were vortexed in heptane to deparaffinize the tissue and were then co-extracted with methanol. The methanol layer was evaporated, and the protein residue was resuspended in 2% SDS/lOOmM ammonium bicarbon-ate/20mM dithiothreitol (DTT), pH 8.5 and heated at 70°C for lh. After tryptic digestion, 123 total confident proteins were identified in the FFPE tissue, compared to 94 proteins identified from the fresh-frozen tissue. Hwang et al. also reported up to a fivefold increase in protein extraction efficiency for samples extracted in a Tris-HCl/2% SDS/1% Triton X-100/1% deoxycholate solution at 94°C for 30 min versus samples extracted in 100 mM ammonium bicarbonate/30% acetonitrile at the same temperature.14... 

Sanchez-Ferrer A, Bru R and Garcia-Carmona F. 1989. Novel procedure for extraction of a latent grape polyphenoloxidase using temperature-induced phase separation in Triton X-114. Plant Physiol... 

Treat sections on the grids with phosphate-buffered saline (PBS) at pH 7.4 containing 0.05% Triton X-100 for 15 min at room temperature. 

Acridinium ester—labeled chemiluminescent probes have been utilized to detect the specific protein-coding transcripts and to distinguish between transcripts that code for the 190-kDa protein and the two closely related 210-kDa proteins. The assay is called the hybridization protection assay (D3). In this assay, RNA isolated from the patient s white blood cells is first amplified by PCR. The amplified product is incubated with the chemiluminescent probe. The unhybridized probe is removed by selective hydrolysis in sodium tetraborate buffer, containing surfactant Triton X-100 at pH 8.5, in an incubation step at 60°C for 6 min. After the sample is cooled to room temperature, the chemiluminescence of the hybridized probe is measured in a luminometer. The procedure is reported to detect one leukemic cell in a population of a million or more normal cells. It is also rapid, requiring less than 30 min. Its reliability has been attested to by correlation with results obtained on karyotypic and Southern blot analysis (D3). 

Pretreatment Solution. 40% [w/v] glucose (tee Note 2), 0.8% [v/v] Triton X-100 (Nacalai) see Note 3-5). Store at room temperature for up to 2 months. Mix well before using. 

Detergents. Under appropriate conditions of pH, ionic strength and temperature, detergents (ionic sodium lauiyl sulphate, sodium deoxycholate, sodium cholate and cetyldiethyl-ammonium bromide, or nonionic Tweens and Tritons), can be used to lyse cells. Detergents may however cause enzyme inactivation and may need to be removed before purification. 

Jain et al. (33) used the microemulsion system Triton X-100/cyclo-hexane/hexanol/water/ammonia to prepare silica nanoparticles with entrapped bioactive macromolecules fluorescein isothiocyanate-dextran (FITC-Dx) (mol. mass 19.6 kD), [125I]tyraminylinulin (mol. mass 5 kD), and horseradish peroxidase (HRP) (mol. mass 40 kD). The biomolecules were first solubilized in the microemulsion, and the alkoxide (TMOS) was then added. To ensure small particle sizes, the reaction was conducted under ice-cold temperatures (in a refrigerator for 72 h). 

Manufacturing processes and equipment are similar to those employed for alcohol ethoxylate preparation. In the absence of steric hindrance, ethylene oxide reacts with both hydrogens of primary amines at relatively low temperatures (90—120°C) without added catalysts (105). When the nitrogen atom is hindered, as it is in the Triton RW products, only one of the amino hydrogens reacts with ethylene oxide. Once this reaction is complete, a basic catalyst is added and ethoxylation proceeds in the manner of the alcohol-based nonionics. In IV-alkyl-l,3-propanediamine, all three amino hydrogens are available for reaction with ethylene oxide. N-Alkyl-1,3-propanediamines are prepared from fatty monoamines and acrylonitrile, followed by reduction of the resulting 3-cyanoethylalkyl amine. 

Solid CH4 on Triton and Terrestrial Methane Hydrate. - 5.4.1 Radicals in Solid CH4 and on Triton. Methane (CH4) and nitrogen are gaseous molecules on Earth but are frozen and solids under the lowest temperature of 37K at Triton, a satellite of Neptune. Craters of solidified CH4 and N2 and black smoke consisting of gaseous N2 and solid CH4 were discovered by the Voyager II a few km above the ground volcanic eruptions in 1989. 

Let the plates come to room temperature and rinse the cover slips twice with PBS. Incubate the cover slips for 5 min with 2 ml Triton X-100 solution, remove, and rinse twice with 2 ml PBS. Remove the PBS and incubate the cover slips for 30 min with 2 ml of filipin assay solution. Remove the filipin assay solution and rinse the cover slips twice with PBS. Remove the PBS and let the cover slips dry at ambient temperature. Mount the cover slips on a microscope slide with one drop of DABCO solution... 

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