Detergentes: Triton X-100, Tween-20 e mais

2013 ◽  
Vol pt3 ◽  
Author(s):  
Mary Johnson
Keyword(s):  
Tween 20 ◽  
1987 ◽  
Vol 65 (1) ◽  
pp. 8-18 ◽  
Author(s):  
Rex K. M. Wong ◽  
Christine P. Nichol ◽  
M. Chandra Sekar ◽  
Basil D. Roufogalis

The efficiency of several nonionic detergents and a homologous series of zwitterionic detergents for the extraction of acetylcholinesterase (EC 3.1.1.7) from bovine erythrocyte membranes was examined. Of the nonionic detergents examined, the polyoxyethylene-based Tweens were the least effective solubilizing agents. Within this series, increasing the length of the saturated fatty acid chain progressively decreased the efficiency of enzyme recovery, while unsaturation in the side chain reversed this trend. In the Lubrol detergents, where the chain length of the alcohol group is variable, an increase in the length of the polyoxyethylene glycol group decreased the recovery of acetylcholinesterase in the solubilized state, without affecting the efficiency of extraction of total erythrocyte protein. As with the other nonionic detergents examined, Triton X-100 and octy1 β-D-glucoside were maximally effective in solubilizing acetylcholinesterase activity at concentrations greater than their respective critical micelle concentrations. In the sulfobetaine (N-alkyldimethylaminopropane sulphonate) zwitterionic detergent series, the longer alkyl chain zwittergents Z 316 and Z 314 were more efficient than the shorter chain length members of the series (Z 310 and Z 312). In contrast to the higher chain length compounds, short chain analogs were maximally effective at or below their critical micelle concentrations. After purification by ion-exchange chromatography and affinity chromatography, the enzyme extracted with the various detergents gave sedimentation coefficients between 6.8S and 7.6S, consistent with a dimeric structure. Acetylcholinesterase could also be efficiently released by 0.2 mM EDTA or 0.5 M NaCl from bovine erythrocyte membranes previously depleted of 70–80% of the membrane lipids by butanol. Nonlinear Arrhenius plots of enzyme activity were found whether acetylcholinesterase was solubilized with Tween 20, Lubrol PX, or Triton X-100. The present work confirms that bovine erythrocyte acetylcholinesterase requires detergents to solubilize it from membranes and that its activity depends on the structure of the amphiphiles used to solubilize the enzyme.


1985 ◽  
Vol 227 (2) ◽  
pp. 405-412 ◽  
Author(s):  
P W Cheng ◽  
W E Wingert ◽  
M R Little ◽  
R Wei

We have characterized a bovine tracheal mucin beta-6-N-acetylglucosaminyltransferase that catalyses the transfer of N-acetylglucosamine from UDP-N-acetylglucosamine to the C-6 of the N-acetylgalactosamine residue of galactosyl-β 1→3-N-acetylgalactosamine. Optimal enzyme activity was obtained between pH 7.5-8.5, at 5mM-MnCl2, and at 0.06-0.08% (v/v) Triton X-100 (or Nonidet P-40), or 0.5-5.0% (v/v) Tween 20. Ba2+, Mg2+ and Ca2+ could partially replace Mn2+, but Co2+, Fe2+, Cd2+ and Zn2+ could not. Sodium dodecyl sulphate, cetylpyridinium chloride, sodium deoxycholate, octyl beta-D-glucoside, digitonin and alkyl alcohols were less effective in enhancing enzyme activity, and dimethyl sulphoxide was ineffective. The apparent Michaelis constants were 1.25 mM for UDP-N-acetylglucosamine, 0.94-3.34 mM for freezing-point-depressing glycoprotein and 0.19 mM for periodate-treated blood-group-A porcine submaxillary mucin. Asialo ovine submaxillary mucin could not serve as the glycosyl acceptor. The structure of the 14C-labelled oligosaccharide obtained by alkaline-borohydride treatment of the product was identified as Gal beta 1→3(Glc-NAc beta 1→6)N-acetylgalactosaminitol by beta-hexosaminidase treatment, gas chromatography-mass spectrometry and 1H-n.m.r. (270 MHz) analysis. The enzyme is important in the regulation of mucin oligosaccharide biosynthesis.


2004 ◽  
Vol 87 (3) ◽  
pp. 657-663 ◽  
Author(s):  
Meenakshi Tripathi ◽  
Subhash K Khanna ◽  
Mukul Das

Abstract A simple method has been developed for the extraction, separation, and determination of synthetic colors in ice cream samples. The process involves the breakdown of emulsion by neutral detergents (Triton X-100 and Tween 20) followed by extraction with petroleum ether for removal of fat. The aqueous colored solution obtained is treated with 5% acetic acid, and the uptake of color is carried out by a wool-dyeing technique. The color is eluted from the wool with 5% ammonia solution, the solution is evaporated to dryness, and the residue is dissolved in 60% ethanol for paper chromatography using trisodium citrate–ammonia–water (2 + 5 + 95, w/v/v) as the mobile phase. The colored spots from the paper chromatogram are cut and eluted with 60% ethanol, and the absorbance is measured at the respective λ maximum corresponding to the Rf value of the appropriate standard. The recoveries of 6 colors, including sunset yellow FCF (SSYFCF), tartrazine, carmoisine, ponceau 4R, brilliant blue FCF (BBFCF), and fast green FCF from spiked samples with either detergent were found to be >90%. However, recoveries of erythrosine were 21 and 65% with Triton X-100 and Tween 20, respectively. Indigo carmine could not be recovered at all because of its fugitive property in 5% ammonia solution, which is used to strip the color from the wool. The sensitivity of the method with the use of Tween 20 is 1 ppm (1 μg/g) for the colors in spiked ice cream samples. With this method, we analyzed samples of 20 branded colored ice cream. The results showed the presence of tartrazine (8.4–43.3 ppm), SSYFCF (23.5–117.6 ppm), carmoisine (traces–53.2 ppm), erythrosine (3.5 ppm), and BBFCF (4.1 ppm) in the ice cream samples. Apart from 2 samples of tuttifruity, all of the ice cream samples showed the presence of permitted synthetic colors below the permissible level of 100 ppm established by the Prevention of Food Adulteration Act of India.


2012 ◽  
Vol 47 ◽  
pp. 62-67 ◽  
Author(s):  
G. Ulloa ◽  
C. Coutens ◽  
M. Sánchez ◽  
J. Sineiro ◽  
A. Rodríguez ◽  
...  

2007 ◽  
Vol 55 (8) ◽  
pp. 805-812 ◽  
Author(s):  
Marija Heffer-Lauc ◽  
Barbara Viljetić ◽  
Katarina Vajn ◽  
Ronald L. Schnaar ◽  
Gordan Lauc

Gangliosides and glycosylphosphatidylinositol (GPI)-anchored proteins contain lipid tails that tether them to the outer side of the cell membrane. This mode of association with the cell membrane enables them to take part in the organization of lipid rafts, but it also permits gangliosides and GPI-anchored proteins to be actively released from one cell and inserted into the membrane of another cell. Recently, we reported that under conditions of lipid raft isolation, Triton X-100 causes significant redistribution of both gangliosides and GPI-anchored proteins. Aiming to find a less disruptive detergent, we evaluated the effects of CHAPS, Saponin, deoxycholic acid, Trappsol, Tween 20, Triton X-100, Brij 96V, Brij 98, and SDS on brain tissue sections. At room temperature, all detergents (1% concentration) extracted significant amounts of both gangliosides and Thy-1. At 4C, the extraction was weaker, but Triton X-100, CHAPS, and deoxycholic acid caused significant redistribution of GD1a and Thy-1 from gray matter into the white matter. Both redistribution and extraction were significantly augmented when sections were incubated with detergents in the presence of primary antibodies. Of the nine tested detergents, none is the ideal choice. However, Brij 96V appears to be able to sufficiently reveal myelin epitopes while causing the least amount of artifacts. This manuscript contains online supplemental material at http://www.jhc.org . Please visit this article online to view these materials. (J Histochem Cytochem 55: 805–812, 2007)


2014 ◽  
Vol 10 (1) ◽  
pp. 110-112
Author(s):  
Б. Энхжаргал ◽  
Жин-Вон Ким
Keyword(s):  
Tween 20 ◽  

Судалгааг Сөүл хотын их сургуулийн Байгалийн ухааны сургуулийн Ургамлын өвчин судлалын лабораторт 28 хоногтой чинжүүний үрслэгийг фитофторын 10мл-ийн 104 концентрацитай зооспорийн суспензээр халдварлуулж өвчний эсрэг 2 төрлийн гадаргуугын идэвхит бодис (Triton X-100, Tween-20)-ыг 1000, 2000, 3000ppm тунгаар тооцож үр дүнг тооцсон.DOI: http://dx.doi.org/10.5564/mjas.v10i1.306


2008 ◽  
Vol 55 (1) ◽  
pp. 135-140 ◽  
Author(s):  
Anna Potocka ◽  
Jan Zimowski

A membrane-bound phospholipid : steryl glucoside acyltransferase from Solanum melongena leaves was partially purified and its specificity and molecular as well as kinetic properties were defined. Among the steryl glycosides tested (e.g. typical plant steryl glucosides, steryl galactosides and cholesteryl xyloside) the highest activity was found with cholesteryl glucoside, but some structurally related compounds such as sito- and stigmasteryl glucoside or galactoside as well as cholesteryl galactoside were also acylated, albeit at lower rates. The investigated enzyme was able to use all classes of phosphoglycerolipids (phosphatidylethanolamine, phosphatidylcholine, phosphatidylserine, phosphatidylinositol, phosphatidylglycerol) as an acyl source for biosynthesis of acylated steryl glucoside. Among them 1,2-dimirystoylphosphatidylic acid appeared to be the best acyl donor. Apart from phosphoglycerolipids, 1,2-diacylglycerols were also used as acyl donor for steryl glucoside acylation, although at a distinctly lower rate. The acyl moiety was transferred from the C-1 position of phospholipid molecule. The investigated acyltransferase activity was stimulated by 2-mercaptoethanol, Triton X-100, 1-monoacylglycerols and inhibited in the presence of divalent cations such as Ca(2+), Mn(2+), Zn(2+) or Co(2+), some lipids (MDGD, ceramide), detergents (Tween 20, 40, 60 and 80, Tyloxapol, sodium deoxycholate) and high ionic strength.


1998 ◽  
Vol 123 (6) ◽  
pp. 987-991 ◽  
Author(s):  
Shiow Y. Wang ◽  
Dean Der-Syh Tzeng

Foliar application of a mixture of methionine (1 mm) and riboflavin (26.6 μm) reduced the severity of powdery mildew [Sphaerotheca macularia (Wallr. ex Fr.) Jacz. f. sp. fragariae] infection in `Earliglow' strawberry (Fragaria × ananassa Duch.) plants. Efficacy of this mixture on controlling powdery mildew infection was enhanced by supplements of copper, iron, and surfactants [sodium dodecyl sulfate (SDS), Triton X-100, Tween-20, or oxyalkylenemethylsiloxane (Silwet L-77)]. Free-radical scavengers (n-propyl gallate, thiourea) and antioxidants (α-tocopherol, β-carotene) reduced the efficacy of this mixture. Plants treated with a mixture of riboflavin (26.6 μm), d,l-methionine (1 mm), copper sulfate pentahydrate (1 mm), and surfactants (SDS or Silwet L-77 at concentrations of 0.05% to 0.1%) showed a decrease in powdery mildew infection. Results of this study suggest that treatment with a mixture of methionine and riboflavin is beneficial to strawberry plants and may serve as an alternative to fungicides for controlling powdery mildew.


2004 ◽  
Vol 16 (2) ◽  
pp. 177 ◽  
Author(s):  
H. Men ◽  
Y. Agca ◽  
S.F. Mullen ◽  
E.S. Critser ◽  
J.K. Critser

Disruption of the actin cytoskeleton is one of the leading causes in low survival of pig embryos after cryopreservation (Dobrinsky et al., 2000 Biol. Reprod. 62, 564–570). In this study, the effect of osmotic stress on cellular actin filament organization in porcine embryos produced in vitro was studied. Excellent quality Day 6 (fertilization=Day 0) porcine blastocysts were randomly exposed to 6 different anisosmotic sucrose solutions (75, 150, 210, 600, 1200, 2400mOsm) for 10min. Embryos were then returned to embryo culture medium (NCSU-23) after washing with NCSU-23, and cultured under 38.5°C, 5% CO2 in air with maximal humidity for them to recover. Blastocysts cultured in NCSU-23 medium (280mOsm) served as a control for embryos with intact actin filament organization. Blastocysts treated with 7.5μgmL−1 cytochalasin-b for 60min served as a control for embryos with F-actin depolymerization. Eighteen hours post-anisosmotic treatments, all blastoysts were fixed in 3.7% paraformaldehyde in PBS for 60min and stored in PBS with 0.1% Triton X-100 and 0.2% sodium azide at 4°C. Staining of actin filaments was performed according to procedures described earlier (Wang et al., 1999 Biol. Reprod. 60, 1020–1028). Embryos were blocked in PBS with 20mgmL−1 BSA and 150mM glycine for 30min. After being washed in PBS with 0.1% Tween 20 for 60min, embryos were stained with 10UmL−1 Alexa Fluor 488 phalloidin in PBS with 0.1% Tween 20 at 38.5°C for 60min, and then washed twice in PBS with 0.1% Tween 20 for 60min each. The status of actin filaments in embryonic cells was examined by confocal microscopy. Integrity of cellular actin filaments was classified as either intact or disrupted according to the distribution within embryonic cells. Blastocysts were then classified according to the status of actin filaments in embryonic cells. Data were analyzed using logistic regression. Results from 7 replicates are displayed in Table 1. There was a significant relationship between osmotic treatment levels and the probability of blastocysts with disrupted cellular actin filaments (P<0.0001). These data support the hypothesis that porcine embryos are very sensitive to osmotic changes. Ongoing experiments will assess the extent of actin disruption required to significantly reduce developmental competence of pig blastocysts. This study was supported by Monsanto Company. Table 1 Cellular actin filament integrity of in vitro produced porcine blastocysts after being treated with sucrose solutions with different osmolalities (mOsm)


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