Nonionic detergents triton

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-... 

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

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. 

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.

Analysis of histones is frequently carried out in the presence of detergents which may bind to them. Binding of the nonionic detergent Triton X-100 to histones from calf thymus can be analysed by gel chromatography. The binding of Triton to histones was abolished if 8 M urea was present in 0.01 M HCl or 0.9 M acetic acid [110]. Subsequently [111], the effects of the cationic detergent cetyltrimethyl ammonium bromide (CTAB), and the anionic detergent sodium dodecyl sulphate (SDS), in GPC and electrophoresis were compared. The results showed that, in the presence of CTAB, the values of the retardation and distribution coefficients were linear in respect to the molecular mass of histones, while in the presence of SDS, deviations from this type were observed. 

In 1951 Kellner et al. reported that administration of the nonionic detergents Triton A-20 or Tween 80 to rabbits and guinea pigs was followed by a sustained hyperlipemia. This observation was later extended to other animal species mice (Hirsch and Kellner, 1956a, b), rats (Friedman and Byers, 1953), and dogs (Scanu et al., 1961). Studies by Friedman and Byers (1953, 1957) and Hirsch and Kellner (1956a, b) led to the hypothesis that the hyperlipemia was secondary to the action of the detergent on the physical and chemical properties of plasma lipoproteins. Experimental support for this hypothesis was provided by the... 

Fresh embryos can be fractionated in a manner similar to frozen embryos. However, significant numbers of unbroken cells are observed if procedures are followed exactly as outlined above to ensure complete breakage, the nonionic detergent, Triton X-100, should be added to initial buffers (Buffer E or A) at a final concentration of 0.5-1.0% (v/v). Drosophila tissue culture cells are useful for many purposes (e.g., radiolabeling of proteins). Nuclei can be prepared from both Kc and Schneider S2 cells using procedures identical with those described above for fresh embryos (see Smith et ai, 1987 Smith and Fisher, 1989 Mans et al, 1995). Cells should be harvested from exponentially growing cultures. 

Protein synthesis and membrane formation in embryonic and postnatal rat liver occurs faster than in the liver of adult individuals. It was demonstrated that an increase in transferase activity coincided with an increase in hydrolase activity. However, the ratio between these two activities was not constant. This ratio increased from 0.8 (day of delivery) to 2.6 (4-7 days after delivery) and subsequently decreased to 1-2 (adult rat). These fluctuations were caused by the binding of enzyme molecules with microsomal membranes, and differences in the expression of catalytic properties were caused by the relative proportions of membrane-bound and free fractions. It was demonstrated with the aid of enzyme extractions with the nonionic detergent Triton X-100 and sodium deoxycholate (Duck-Chong and Poliak, 1973 Fig. 38). There are bacterial mutants with reduced esterase activity. In some of them esterase activity is reduced because of low levels of enzyme synthesis. Other mutants have low esterase activity due to the weakening of the association of the membrane-enzyme compound (Frehel et al., 1974). 

FIGURE 3.1 Some examples of common surfactant molecules, from top anionic surfactant SDS (sodium dodecyl sulfate, also commonly known as sodium lauryl sulfate) cationic surfactants CTAB (hexadecyltrimethylammonium bromide), DDAB (didecyidimethylammonium bromide) commercial nonionic detergent Triton X-100 and zwitterionic lipid DOPC (1,2-dioleoyl-sn-glycero-3-phosphocholine). 

Cell fractionation procedures were fundamental to the biochemical identification of steroid and thyroid hormone receptors in brain as well as in other tissues. Isolation of highly purified cell nuclei from small amounts of tissue from discrete brain regions generally is accomplished with the aid of a nonionic detergent, such as Triton X-100 [7],... 

An abbreviation used for Triton X-100, a nonionic detergent that is used to solubilize membrane-bound proteins. 

The SDS-containing solution is supplemented with an excess of a nonionic detergent, e.g., Triton X-100. The electrode buffer is a buffer with low ionic strength, without SDS, and pH nearby the pi of the protein of interest. During electroelution SDS migrates to the anode. 

Cationic detergents such as cetyl-trimethylammonium chloride (9.63) are more effective than anionic soaps such as sodium dodecylsolfonate (9.64, SDS). Nonionic detergents such as Triton X-100 (9.65, octoxynol or (polyethylene glycol)jo-... 

Several of the synthetic detergents used for dissolving membranes and solubilizing integral membrane proteins. Triton X-100 and octylglucoside are nonionic detergents cetyltrimethylammonium bromide and sodium dodecylsulfate (SDS) are ionic. SDS is also an effective denaturant of proteins and is used in polyacrylamide-gel electrophoresis (see chapter 6). 

Wie, S.I. and B.D. Hammock. 1982. The use of enzyme-linked immunosorbent assays (ELISA) for the determination of Triton X nonionic detergents. Anal. Biochem. 125 168-176. 

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