Pipes, solution preparation

After primary fixation the algal and root samples were rinsed in 0.1 M PIPES buffer, pH 7.0, then twice in 0.1 M sodium cacodylate buffer, pH 6.8, and post-fixed in 1% osmium tetroxide in 0.1 M cacodylate buffer, pH 6.8, overnight at 4°C. The osmium solution prepared for the root samples was added with 0.7% potassium ferricyanide in order to improve osmium penetration in the root tissues. This was particularly necessary for the 2 h control roots and roots from RO-treated seed, due to their relatively low water content. The replacement of PIPES buffer with cacodylate buffer before osmication was necessary, as PIPES reacts with osmium, producing a dark precipitate. 

Because lime is a weak base, a solution of pure CaCb is acidic. The commercial form, however, contains a small amount of unreacted lime and produces solutions that are slightly alkaline. Solutions therefore are noncorrosive unless aerated. Carbon steel batch tanks or solution preparation tanks with their frequently varying levels should be lined or painted to prevent corrosion. FRP is also an acceptable material, provided that tanks are designed for the high specific gravity of the material. Carbon steel is also a satisfactory material for pumps and piping. PVC is usefiil up to a temperature of about 70°C. 

In a 1,000 ml three-necked flask equipped with a stirrer, a dropping funnel and a silica gel guard pipe, 46.7 g hydroxylamine hydrochloride are dissolved cold in 480 ml methanol. Separately a solution of 56.1 g KOH in 280 ml methanol is prepared, heated to 30°C and edmixed, drop-wise under stirring to the hydroxylamine solution. All successive temperature increases dur-... 

Zinc Analysis. For EMI of zinc accumulation of Ps. 244, the ligand salicylaldehyde-2-quinolylhydrazone (SAQH) was used. The SAQH was prepared by refluxing sallcylaldehyde (Aldrich) with 2-hydrazlnoqulnollne (Kodak) as described by Sommer al. (12). In methodology analogous to the tin work described above, known quantities of biomass were exposed to 1.0-30.0 ppm aqueous solutions of ZnBtj. Cell samples were periodically withdrawn from the zinc solution in attempts to measure the kinetics of accumulation of the metal. For EMI analysis cells were suspended in 0.20 mL PIPES and were then treated with 0.10 mL of 3.0 x 10 M SAQH in EtOH. The cell samples were then rewashed and resuspended in 0.30 mL PIPES for EHI analysis (A, =366 nm, A, =498 nm). 

Prepare a QD solution in 25 mM PIPES buffer, pH 7.0 (reaction buffer), at a concentration of lOOpg/ml. 

Hydroxide. Potassium hydroxide, [CAS 1310-58-3]. caustic potash, potassium hydrate, KOH, white solid, soluble, mp 380 C, formed (1) by reaction of potassium carbonate and calcium hydroxide in H2O, and then separation of the solution and evaporation. (2) by electrolysis of potassium chloride under the proper conditions, and evaporation. Used in the preparation of potassium salts f 1) in solution, and (2) upon fusion. Also used 111 the manufacture of (3) soaps, (4) drugs. (5) dyes, (6) alkaline batteries, (7) adhesives, (8) fertilizers, (9) alkylates, (10) for purifying industrial gases, (11) for scrubbing out traces of hydrofluoric add in processing equipment, (12) as a drain-pipe cleaner, and (13) in asphalt emulsions. 

As for Basic Protocol 1, the LOX enzyme is prepared according to the Support Protocol. However, more care should be taken to reduce turbidity and other UV-absorbing materials caused by suspended lipid and pigments, especially when low activity is expected (requiring more enzyme addition). Triton X-100 is a UV absorber, and it causes more lipid and pigments to be suspended. Partial purification may be necessary. HEPES and PIPES buffers are not used in this method because of excessive absorption at 234 nm (0.5 to 0.6 absorbance for 0.1 M solutions) MES is useful with limitations (0.2 absorbance for 0.1 M solution). 

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