Tri-bromophenol

Also obtained (poor yield) by alkaline condensation of benzotrichloride with 2,4,6-tri-bromophenol in 30% aqueous sodium hydroxide solution at 90-95° (2%) [127]. 

Red algae (Rhodophyceae) synthesize a substantial range of halogenated compounds including mono- and dihydroxy Ce—Ci, Cg—C2 and Ce—C3 phenols containing one or two bromine atoms (Fenical 1975). 2,4,6-Tri-bromophenol (Figure 7.54) predominates and the total bromophenols content ranges from 8 to 180 p,g/kg. 

Immediately after irradiation, stop the reaction by the addition of 7 pi of 4 X SDS electrophoresis loading buffer or the equivalent (with a high concentration of reducing agent present) 0.2M Tris, 8 percent SDS, 2.88M P-mercaptoethanol, 40 percent glycerol, 0.4 percent xylene cyanol, 0.4 percent bromophenol blue. Heat the sample at 95°C for 5 minutes and analyze the complexes formed by electrophoresis. 

Sodium dodecyl sulfate (SDS) sample buffer stock solution Dissolve 1.51 g Tris base to 15 ml of distilled water, and then add 25 ml of 50% glycerol into the mixture. Stir to dissolve and then adjust pH to 6.8 with HCl. Dissolve 5 g of SDS in this buffer and make up to 50 ml with distilled water. Dissolve 0.01 g of bromophenol blue and stir overnight. 

The remaining dabsyl chloride is not removed, since it reacts with Tris of Soln. C to a dye acting as tracking dye like bromophenol blue. 

SDS-PAGE loading buffer (4x). 200 mM Tris-HCl, pH 6.8, 8% SDS, 32% glycerol, 0.008% bromophenol blue, 8% 2-mercapto ethanol Store at room temperature. 

Electrophoresis apparatus for vertical slab gels and power supply Sample application buffer Tris, glycerol, bromophenol blue, pH 6.8 Protein solution buffer Tris, pH 6.8... 

Gel-loading buffer, 0.04 M Tris-acetate containing 50% glycerol and 0.25% bromophenol blue tracking dye, pH 8.0. 

Electrophorese a 5 m1 sample of the above digest in parallel with 4m1 of the untreated RFI solution on a 1% agarose gel in tris-borate buffer (lOOmM-Tris base, lOOmM-boric acid, 2 mM-EDTA). Dilute each sample into 15 m1 H20 and add 3m1 loading buffer (3% bromophenol blue in 0.1M-EDTA, pH 8.3, 50%... 

After the stacking gel has polymerized for at least 30 min, the sample comb is removed and the upper and lower reservoir chambers are filled with Electrode Buffer (25 mM Tris-HCl, 192 mM glycine, 0.1% SDS, pH 8.3). The samples are prepared by mixing four parts of sample with one part of 5X Sample Buffer (175 mM Tris-HCl, pH 6.8, 11% SDS, 0.14% bromophenol blue, 55% glycerol, +2 M DTT). The samples are heated at 95°-100° C for 2-5 min. Each sample well is rinsed with Electrode Buffer before applying the samples to individual wells. The gel s electrodes are then connected to the power supply and the gels are run at a constant current of 40 mA per gel until the tracking dye reaches the bottom of the gel ( 4 hr). 

X SDS/EDTA buffer—Dissolve 3.51 g of Tris HC1 and 0.335 g of Tris free base in 50 ml of 0.5 M EDTA, pH 8.0. Adjust the pH to 7.0 with dilute NaOH or HC1 if necessary. Add 30 ml of glycerol, 10 ml of /3-mercaptoethanol, and 10 ml of distilled water. Dissolve (avoid foaming) 8 g of SDS into this solution, as well as 4 mg of bromophenol blue. Store the solution tightly capped at room temperature. 

Prepare a bromophenol blue solution by dissolving 10 mg dye in 5 ml ethanol. Use a vortex mixer to make certain all of the dye dissolves. Add, dropwise, IM Tris-acetate buffer (pH 7.0) until the solution turns intensely blue. 

Fig. 4. A-D. Resonance Raman spectra of pyrocatechase complexed with o-fluorophenol (A) o-chlorophenol (B) o-bromophenol (C) o-chlorophenol-4,6-d2 (D). Conditions 647.1 nm excitation, 150-200 mW power, 4 cm-1 slit width, 40-50 mg/mL of enzyme in Tris-acetate pH 8.5 buffer, 4 °C sample temperature. SO - was used as internal standard...

X loading buffer 50 mM Tris-borate, 1 mMEDTA, 0.1% xylene cyanol, 0.1% bromophenol blue, in 100% formamide. 

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) buffer 100 mM Tris-HCl, pH 6.8,4% 2-mercaptoethanol, 4% SDS, 0.2% bromophenol blue, 20% glycerol. 

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