For ascorbate peroxidase assay extraction 241 buffer was supplemented with 1.0 mM ascorbic acid. The homogenate was centrifuged at 242 15,000×g for 15 min at 40C, and the supernatant was used as a crude enzyme extract. 243 Spectrophotometric determinations were performed using UV visible spectrophotometer 244 (UV-1700, Shimadju, Japan). 245 2.11.2. Estimation of superoxide dismutase (SOD) activity 246 SOD activity was estimated by its ability to catalyse NBT to formazan at 560nm 247 according to the method of Beyer and Fridovich (40).
Experimental Methods: 1. SYNTHESIS OF 4-BENZOYL BUTYRIC ACID METHYL ESTER Materials required * 5 oxopentanoic acid : 2 gm (Aldrich) * Methanol : 50 ml * Acetic Acid (Rankem) Procedure: * 2 grams of 5 oxopentanoic acid was weighed and placed in a round bottom flask and then to it 50 ml of methanol was added. It was placed on a hot plate and the temperature was increased to 50 degrees under ambient air conditions. * To the RB, 2 ml of acetic acid was added and then by attaching a condenser the entire reaction was put on refluxing at 70 degrees Celsius in an oil bath. * For work up: * The reaction media was concentrated till about 10 ml and then dry silica gel was added.
Figure XD shows a record of 30 s long of the patch currents from an oxaliplatin treated neuron in the presence of 1 M of icilin at Vm= 40 mV (upper trace) in which at least three channels were active. Moreover, the red open circles of figure XE represent the single channel current amplitudes recorded at different membrane potentials. In this condition, the fitted regression line yields a and a Vr respectively of 1 00 pS and of 0
Second part of blood was left to coagulate then centrifuged at 3000 rpm for 15 minutes to obtain serum to estimate some biochemical parameters. Serum insulin and leptin were estimated according to the methods of Wilson and Miles, (1977) and Palacio et al. (2002) respectively. (TG) and high density lipoprotein cholesterol (HDL-c) were determined by using enzymatic colorimetric methods described by Kostener et al. (1977).
The tube was placed back in incubation for 96 more hours to observe any more positives. 2.10 Catalase Test A trypticase soy agar plate was used and after incubation, four drops of 3% Hydrogen Peroxide was added to the plate to flow over the bacterial growth. A presence of bubbling was observed. 2.11 Starch Hydrolysis A starch agar plate was inoculated with a streak of the unknown bacteria and then incubated. On the second day of incubation, the plate was removed from the incubator and placed over a hot plate heating Iodine solids.
Firstly, 0.3 g Fe3O4 was ultrasonically dispersed into 150 ml solvent (Vmethanol/Vwater=1/1) for 20 min to obtain a uniform suspension. A 100 ml alkaline solution (0.32 g Na2CO3, 0.48 g NaOH) was added dropwise into the suspension until pH ca. 10.0 and kept for 5 min, then another 100 ml salt solution (1.02 g Mg(NO3)2⋅6H2O and 1.2 g Al(NO3)3⋅9H2O) and above alkaline solution were simultaneously added maintaining the pH in 9.5-10.0. The resultant was stirred for 5 min followed separation by using a magnet and thoroughly washing with deionized water and alcohol and dried at 60 °C overnight giving product Fe-Mg/Al
The resulting reaction between the two chemicals is such that for each gram of A, 4 grams of B is used. It is observed that 30 grams of the compound C is formed in 10 minutes Determine the amount of C at time t if the rate of the reaction is proportional to the amounts of A and B remaining and if initially there are 50 grams of A and 32 grams of B. How much of the compound C is present at 15 minutes? Interpret the solution as t →∞ ? In this example we will use the rate equation .
2. Prepare the electrophoresis tank by filling the tank with 900ml of TEB buffer wicks are cut from Grade Number 3 chromatography paper and were placed along the 22 cm long bridges in the tank. 3. The cellulose acetate membranes are cut in 40xl00 mm each and soaked in TEB buffer for 5 minutes. 5 strips are plotted then placed on the electrophoresis tanks.
The resolution was kept at 4 cm-1 and scanning time was fixed at38 Sec. A total number of 10 scans were carried out on each sample. 3. RESULTS ANDDISCUSSION: Fig.1. shows FTIR spectra of human blood serum treated with Urea of concentrations 100, 200, 300, 400, 500 mg/dl.
32 100 μL of afore-prepared sample solution and the mixed reference standard were diluted 100 times with ethyl acetate. 50 μL of these dilution solutions were separated on the TLC plate coated with SNISG. The plate was developed with petroleum ether: ethyl acetate (4:1) and the movement of solvent was usually controlled at 1 cm from the upper edge. After completion, the plate was dried until no solvent smell remained. It was sprayed with an ethanol solution containing 10% sulfuric acid, and heated at an infra-red drier until obvious color came up, as shown in Fig.2 (B.ab).
15 mL of Solution A and B were mixed together to form solution F. Eight cuvettes were labeled distinctly as 1a, 2a, 3a, 4a, 1b, 2b, 3b, 4b, where “a” cuvettes were used for the concentration experiment and “b” cuvettes were used for the temperature experiment. Cuvette 1, the blank tube was prepared and the spectrophotometer was set to 405 nm. The enzyme was added, upon being ready to start the experiment, to tube 1 which then became tube “1a.” 3 mL of solution F was added to each cuvette, both “a” and “b.” The “b” cuvettes were then placed in their specific temperatures, 1b in the fridge, 2b in room temperature, 3b in a 32 degrees Celsius water bath and 4b in a 60 degree water bath. The temperature was recorded using a thermometer that was placed in the surroundings of the tube. The cuvettes were retrieved from their respected conditions.
After adding 20 microliters of proteinase K to the 1.5 ml tube, the sample was vortexed for 30 seconds to disrupt the pellet. The sample was then incubated at 56°C to lyse the tissue. The sample was checked every fifteen minutes and vortexed between each checking for an hour and a half until the tissue was completely lysed. The tissue sample was then again vortexed. Next 200 microliters of buffer AL was added and