Detergent Triton

Hg ", Zn ", Cd " ) light emission is shifted to the red (610—615 nm). In vitro a flash of light is produced (< 1 s) that decays rapidly. Glow-type emission is obtained ia the presence of detergents (Triton X-100), polymers (PEG 6000), coen2yme A, inorganic pyrophosphate, and cytidine nucleotides (206,207). 

Since it might be possible that the perturbation of membrane directly stimulated the NADPH-oxidase located on the cell membrane, which is the enzyme for the production of superoxide [24], the possibility was examined by the assay using detergent (Triton X-100) instead of polymers. At 0.001% of Triton X-100, no stimulation of superoxide release from DMSO-differentiated HL-60 cells was observed. At 0.01% of Triton X-100, a... 

Vermelho AB, Pereira MC, Meirelles M. Use ofthe detergent Triton in biochemical and biological research a minireview. Ciencia Cultura 40 45-51. 

Since hematin inhibits Taq polymerase, it is absolutely essential to eliminate red blood cell contamination. Selective lysis of red blood cells can be accomplished with a buffer mixture consisting of 155 mM ammonium chloride, 10 mM potassium bicarbonate, and 0.1 mM EDTA adjusted to pH 7.4. Alternatively, the cytoplasmic membrane of all cells can be dissolved with a buffer mixture containing the non-ionic detergent Triton-X 100, leaving behind nuclei of white blood cells from which DNA can be extracted. However, this technique will result in the loss of cytoplasmic DNA to the supernatant, and hence will not be able to extract mitochondrial DNA (B11). 

Polyoxyethylene nonionic detergents (Triton X-100, Nonidet P-40, Tween-20, Brij 35, etc.) are used most often in immunochemical techniques because they generally do not denature proteins. Detergents act by intercalating into phos-... 

Rubredoxin is an electron-transfer protein with an Fe(IlI)/Fe(lI) redox couple at -0.31 V (SCE) in water (20). Our peptide model, [Fe( Cys-Pro-Leu-Cys-OMe)2] (Z = benzyloxycarbonyl) (21) exhibits its Fe(lll)/Fe(ll) redox couple at -0.50 V (SCE) in Mc2SO (9). This is similar to the value observed for the native protein when the difference of the solvent is taken into account. When the model complex is solubilized in water by formation of micelles with addition of the non-ionic detergent, Triton X-KX), we also observed a quasi-reversible redox couple at -0.37 V (SCE) (5). The Fe(lll) complexes of Cys-X-Y-Cys peptides also exhibit a characteristic MCD band at 350 nm due to ligand-to-metal charge transfer which has also been found in oxidized rubredoxin (4). 

This is commonly done by adding a nonionic detergent, Triton X-100, which does not react with the enzyme. 

A new detergent, Triton-101, in association with p-xylene is used for the suspension of 10 ml. of water with a Y value of approximately 0.5 nCi/liter. The application of a new instrumental technique with three photomultipliers decreases further the Y value of both mixtures. A Teflon cylinder with a volume of 250-300 ml. is used as sample container for low level counting with a Y value of approximately 0.2 nCi/liter. Selected results of samples collected during 1967 are reported, and the radiation dose to the population of the United States from tritium is estimated to be approximately 0.2 mrem./year. 

We found the 31p-NMR chemical shift of monomeric dihexanoyl PC increases upon the addition of the nonionic detergent Triton X-100. This phenomenon was used to quantitate the solubilization of this phospholipid by the detergent micelles as a function of detergent concentration using a simple phase separation model ( 5). Similar studies were carried out on dibutyryl PC. At a phospholipid concentration of 7 mM and 56 mM detergent, 85% of the dihexanoyl PC, but only 3% of the dibutyryl PC was incorporated into the micelles. 

In mammals, red blood cells do not contain DNA since they are devoid of nucleus. The hemoglobin in them can get adsorbed to DNA if it is present during the isolation procedure. Hence for the isolation of DNA from blood, red blood cells are first removed either by Ficoll/Hypaque gradient centrifugation, or lysed by the detergent Triton X-100 followed by recovery of nuclei of white blood cells, which carry DNA. 

Figure 18 Effect of nonionic detergent (Triton X-100) on benzydamine N-oxygenation (FMO) and N-demethylation (CYP) by human liver microsomes. Benzydamine (500 pM) was incubated with pooled human liver microsomes (1.0 mg protein/mL) in tricine buffer (50 mM, pH 8.5 at 37°C) with or without Triton X-100 [1% (v/v)]. Reactions were initiated by the addition of an NADPH-generating system and stopped after 10 minute by the addition of an equal volume (500 pL) of methanol. Precipitated protein was removed by centrifugation, and an aliquot (25 pL) of the supernatant fraction was analyzed by HPLC with fluorescence detection. Abbreviations FMO, flavin monooxygenase CYP, cytochrome P450.

Potentiometric reductive titration, using both fresh thylakoids and PS II particles previously oxidized with ferricyanide, has revealed that the LP couple exhibits a constant midpoint redox potential ( o- +° 12 v ) above pH 7.6, but becomes pH-dependent below this pH, with a slope of about -60 mV pH pH unit, whereas the HP couple is pH-independent in the pH range between 6.5 an 8.5 ( o-+0 36 V) in general, cytochrome b-559 exhibits potential values about 40 mV lower in thylakoids than in PS II particles. After mild heating of the fresh preparations or treatment with the detergent Triton X-100, the HP couple is converted into the LP couple, which preserves its characteristic pH-dependence. In contrast, in the presence of the uncoupler CCCP, the HP couple is also converted into the LP couple, but the pH-dependence proper to the latter is now lost. 

It should be noted that, in spite of the solubilities of most yeast exopolyphosphatases, the detergent Triton X-100 was the best stabilizer of these enzymes during purification and storage (Andreeva et al, 1990 Andreeva et al, 1998a,b). 

Capturing conditions broken cell suspension at a concentration of 3.6% solids in 30 mM ammonium acetate pH 5.5 the suspension contains 1% detergent Triton X-100 Flow rate 300 cm/hr Wash 30 m M ammonium acetate pH 5.5... 

The buffer itself should not effectively compete with a protein for coordination to the metal ligand. Sodium phosphate or sodium acetate are recommended buffers (depending on the pH choice) and the presence of EDTA or sodium citrate should be avoided. The presence of detergents (Triton X-100, Tween-20, urea, etc.) in the buffer does not normally affect the adsorption of proteins [4]. 

All main bile salts (glyco- and taurocholate, deoxycholate and chenodeoxycho-late) have the same effects on HGL activity which increases up to a bile salt concentration of around 3 mM for a single species and 4 mM for a mixture of bile salts. However, it is worth noting that synthetic detergents (Triton X-100, Tween, benzalkonium chloride) that dramatically decrease surface tension (<8mN/m), actually inhibit HGL. 

LPVernon, ER Shaw and B Ke A photochemicaiiy active particte derived from chtoroptasts by the action of the detergent Triton X-100. J Bioi Chem 241 4101-4109... 

---