Triton-XlOO

Micelles (CTAB, SDS, Triton-XlOO) Aqueous surface Fluorescence (HC, AC) 32 448... 

Penetration of the biomembrane is undoubtedly essential for most membrane activity. Araki and Rifkind (13) obtained esr spectra of stearic acid spin labelled erythrocyte membranes in the presence of diverse compounds including Triton XlOO, chlorpromazine and glutaraldehyde. The two surfactants chlorpromazine and Triton XlOO both increase the rate of haemolysis and are shown to increase membrane fluidity. Glutaraldehyde as expected decreases fluidity and decreases the rate of haemolysis. 

The SDS-free PAGE system described by Panyim and Chalkley is suitable especially for small basic proteins (e.g., histones). Additives such as non-ionic detergents as Triton XlOO [0.05-0.1% (w/v)] may additionally increase the resolution. 

Prepare a 9% polyacrylamide separating gel containing 100 /zl/ml gelatin with 3% (wt/vol) stacking gel. Apply samples in sample buffer, without prior heating or reduction. Remove SDS from the gel by incubation in 2.5% (vol/vol) Triton XlOO for 30 min. Incubate the gel at 37° C overnight in enzyme buffer. Stain the gel for 30 min in staining solution and de-stain in the same solution without dye. 

Eppendorf centrifuge for 30 min at maximum speed (14000 r.p.m). The crude membrane fraction (pellet) is resuspended by brief sonication in 150 1 HS supplemented with 1 % Triton XlOO and centrifuged again at 4 C for 30 min at 14000 r.p.m. The proteins (including VAMPs) extracted from the membranes during the detergent treatment are found in the supernatant. 

NBD-PC Fluorescence Polarization Method The NBD-PC method, also called FPol, is described in detail on the Evolve site that accompanies this book. The specimen is amniotic fluid that has been centrifuged at 400 Xg for 2 minutes. Results are expressed using artificial units of mil-lipolarization (P x 1000, mP). The analytical measurement range is 150 to 350 mP. No calibrators are used, but use of a Triton XlOO control assures that the temperature is correct and that the dye is not degraded. Comparison of the TDx FLM II and the NBD-PC assay reveals, as expected, a strong nonlinear, inverse correlation ... 

Attention was subsequently directed to the structure of PS I and characterization of the protein structure of photosystem I was initiated hy using a so-called native PS-I complex in which the in vivo structural, functional and spectroscopic characteristics of photosystem I are retained. Such a native PS-I complex was first prepared by Mullet, Burke and Arntzen, who solubilized the thylakoid membrane with a low concentration of Triton XlOO in the absence of salts. The complex was free of cytochrome bffsmd, judged by fluorescence measurements, was also free of certain chlorophyll proteins while retaining the native character of photosystem I. 

CuS has been synthesised by using two different types of w/o microemulsions [29], namely (i) water/cyclohexanone/AOT and (ii) water/cyclohexanone/Triton XlOO/z-propanol. The shape of the CuS obtained from the second was found to be spherical particularly at lower to values, while at higher values (to = 14) the shape was not completely spherical. The size of the nanoparticles increased from 38 to 95 nm with an increase in to from 2 to 14. It can be seen from Fig. 6.3 that at to = 2, the CuS shapes are spherical. 

Permeabilizers Triton-XlOO, Tween 20, saponin, 1-lysophos-phatidylcholine, n -octyl-beta-d-glucopyranoside. 

Nitrocellulose Hi-Flow plus membrane is often used widely. Nitrocellulose membranes are completely neutral, and their binding properties are independent of the pH of the immobilization solution (although pH can have an effect on both the solubility and immobilization efficiency of a particular protein). The immobilization buffer with pH 7.0-7.2 is chosen. Sometimes the surfactants and detergents such as Tween-20 and Triton-XlOO in very low concentration are added in the buffer to reduce the background and nonspecific binding. 

In a next step we tried to dock the physiological substrate, protoporphyrinogen IX, to the INH and Triton-XlOO cleaned enzyme. This attempt failed, although the maximum overlap volume and the dash factors were modified in such a way that the narrow binding pocket was apparently relaxed and, subsequently, even the co-factor FAD was (non-physiologically) removed. Only the product of the... 

Spans (Span 20, Span 40, Span 60, Span 80), Tweens (Tween 20, Tween 80), cyclodextrins, lecithin, sodium dodecyl sulfate (sodium lauryl sulfate), polyethoxylated alcohols, cetyltrimethylammonium bromide (CTAB), Triton XlOO, Brij 721, bile salts (sodium deoxycholate, sodium cholate), methylbenzethonium chloride (Hyamine), and poloxamer 407. 

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