Acid blue membrane

The major stmctural feature of the HAz chain (blue in Figure 5.20) is a hairpin loop of two a helices packed together. The second a helix is 50 amino acids long and reaches back 76 A toward the membrane. At the bottom of the stem there is a i sheet of five antiparallel strands. The central i strand is from HAi, and this is flanked on both sides by hairpin loops from HAz. About 20 residues at the amino terminal end of HAz are associated with the activity by which the vims penetrates the host cell membrane to initiate infection. This region, which is quite hydrophobic, is called the fusion peptide. 

Several compounds that inhibit vesicular glutamate transport have been identified These include the dyes Evans Blue and Rose Bengal. In addition, the stilbene derivative 4,4 -diisothiocyanatostilbene-2,2 -disulfonic acid (DEDS), a compound commonly used as a specific inhibitor of anion channels, inhibits vesicular glutamate transport. Most known inhibitors have limited use as they are membrane impermeant, with the exception of Rose Bengal. 

Figure 52-3. Diagrammatic representation of the major proteins of the membrane of the human red blood cell separated by SDS-PAGE. The bands detected by staining with Coomassie blue are shown in the two left-hand channels, and the glycoproteins detected by staining with periodic acid-Schiff (PAS) reagent are shown in the right-hand channel. (Reproduced, with permission, from Beck WS, Tepper Ri Hemolytic anemias iii membrane disorders, in Hematology, 5th ed. Beck WS [editor]. The MiT Press, 1991.)...

The quality of RNA and transfer can be evaluated by methylene blue staining. This is done by immersing the membrane in 5% acetic acid for 5 min and then in 0.1% methylene blue in 5% acetic acid for 5 min. The membrane is then washed in water, and distinct bands of the 25S (3400 nts), 18S (1800 nts) rRNA, and RNA marker bands should appear. Mark the positions of these with a pencil because the methylene blue staining will disappear during the hybridization. 

FIGURE 7-7 Structure of the P0 glycoprotein protomer. (A) In this ribbon diagram of the extracellular domain of P0, each P strand is labeled with a letter and two antiparallel P sheets are formed. The disulfide bridge is indicated in dark orange and a hypothetical path for disordered amino acids 103-106 is shown in black in the FG loop. (B) Lattice formation by P0. A view of the intraperiod line, or extracellular apposition, of myelin in the PNS. The orange tetramer sets emanate from one bilayer and the blue tetramer interacts with all four of them. This is a view perpendicular to the plane of the myelin membrane. 

FIGURE 50-3 Amino acid sequence conservation across mammalian odorant receptors. ORs pass through the plasma membrane (blue box) seven times, with the AT-terminus located extracellularly and the C-terminus intracellularly. The degree of conservation of each amino acid in this consensus OR is indicated by a colored ball, with dark blue being most highly conserved and red most highly variable. Modified from [5], with permission. 

Mammalian COX (the illustration shows the enzyme from bovine heart) is a dimer that has two identical subunits with masses of 204 kDa each. Only one subunit is shown in detail here the other is indicated by gray lines. Each subunit consists of 13 different polypeptides, which all span the inner mitochondrial membrane. Only polypeptides I (light blue) and II (dark blue) and the linked cofactors are involved in electron transport. The other chains, which are differently expressed in the different organs, probably have regulatory functions. The two heme groups, heme a (orange) and heme ai (red) are bound in polypeptide 1. The copper center Cua consists of two copper ions (green), which are coordinated by amino acid residues in polypeptide II. The second copper (Cub) is located in polypeptide I near heme... 

Reflectance measurements provided an excellent means for building an ammonium ion sensor involving immobilization of a colorimetric acid-base indicator in the flow-cell depicted schematically in Fig. 3.38.C. The cell was furnished with a microporous PTFE membrane supported on the inner surface of the light window. The detection limit achieved was found to depend on the constant of the immobilized acid-base indicator used it was lO M for /7-Xylenol Blue (pAT, = 2.0). The response time was related to the ammonium ion concentration and ranged from 1 to 60 min. The sensor remained stable for over 6 months and was used to determine the analyte in real samples consisting of purified waste water, which was taken from a tank where the water was collected for release into the mimicipal waste water treatment plant. Since no significant interference fi-om acid compounds such as carbon dioxide or acetic acid was encountered, the sensor proved to be applicable to real samples after pH adjustment. The ammonium concentrations provided by the sensor were consistent with those obtained by ion chromatography, a spectrophotometric assay and an ammonia-selective electrode [269]. 

The sensing microzone of the flow-through sensor depicted in Fig. 5.9.B1 integrates gas-diffusion and detection with two analytical reactions [28], viz. (a) the urease-catalysed formation of ammonium ion by hydrolysis of urea (the analyte), which takes places on a hydrophilic enzyme membrane in contact with the sample-donor stream, which contains a gel where the enzyme is covalently bound and (b) an acid-b reaction that takes place at the microzone on the other side of the diffusion membrane and involves Bromothymol Blue as indicator. This is a sandwich-type sensor including a hydrophilic and a hydrophobic membrane across which the sample stream is circulated —whence it is formally similar to some enzyme electrodes. Since the enzymatic conversion of the analyte must be as efficient as possible, deteetion (based on fibre optics) is performed after the donor and acceptor streams have passed through the sensor. Unlike the previous sensor (Fig. 5.9.A), this does not rely on the wall-jet approach in addition, each stream has its own outlet and the system includes two sensing microzones... 

Dye oxidation (e.g., tetrazolium reductase activity with 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide, MTT 2-[4-iodophenyl]-3-[4-nitrophenyl]-5-[2,4-disulfophenyl]-2H tetrazolium monosodium salt, WST-1 3- (4,5 -carboxymethoxyphenyl) -2-(4-sulfophenyl)-2 H-tetra-zolium, MTS 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide inner salt, XTT 2,2 -di-p-nitrophenyl-5,5 -diphenyl-3,3 -(3,3 -dimethoxy-4,4 -diphe-nylenej-ditetrazolium chloride, NET), Alamar blue assays, ATP concentration (e.g., luciferase assay), oxygen consumption (e.g., oxygen electrodes, phosphorescent oxygen-sensitive dyes), mitochondrial protein and nucleic acid synthesis mitochondrial mass (e.g., mitotracker dyes) mitochondrial membrane potential (e.g., tetramethylrho-damine methyl ester, TMRM tetramethylrhodamine ethyl ester, TMRE)... 

Proteins blotted on PVDF membranes are stainable with Coomassie Brilliant Blue R250, but a relative intense background remains, which does not influence, for example, amino acid sequence analysis. 

Mild, selective oxidation of the sialic acid side-chain, followed by the addition of bisulfite and Toluidine Blue has been used for topo-optical staining of sialoglycoconjugates in erythrocyte and lymphocyte membranes. This method, which is based on a polarization-microscope technique, pennits not only specific, and very sensitive, light-microscope staining of sialic acid residues on cell surfaces but also the study of the spatial arrangement of sialoglycoconjugate molecules on cell surfaces.192,197-198... 

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