User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/11/13
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==ADA Kinetic Assay Preparations== | ==ADA Kinetic Assay Preparations== | ||
* A weight of 0.6702 g of sodium phosphate dibasic was dissolved in 50 mL of water to obtain a molarity of 0.05 M. The pH of the solution was adjusted to pH 7.4. | * A weight of 0.6702 g of sodium phosphate dibasic was dissolved in 50 mL of water to obtain a molarity of 0.05 M. The pH of the solution was adjusted to pH 7.4. | ||
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.050 L of water × <math>\frac{0.05 mol}{1L}</math> = .0025 mol of Na<sub>2</sub>HPO<sub>4</sub> × <math>\frac{268.07 g}{1 mol}</math> = 0.6702 g | .050 L of water × <math>\frac{0.05 mol}{1L}</math> = .0025 mol of Na<sub>2</sub>HPO<sub>4</sub> × <math>\frac{268.07 g}{1 mol}</math> = 0.6702 g | ||
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* The pH was was adjusted to 7.4 by the addition of 2 drops of 12 M HCl. | * The pH was was adjusted to 7.4 by the addition of 2 drops of 12 M HCl. | ||
* To obtain 1 mM inosine, 1.5 mg of the solid was dissolved in 1 mL buffer. This was further diluted by collecting 89.3 µL of the 1.5 mg/ml inosine into 5 mL of the sodium phosphate buffer. | * To obtain 1 mM inosine, 1.5 mg of the solid was dissolved in 1 mL buffer. This was further diluted by collecting 89.3 µL of the 1.5 mg/ml inosine into 5 mL of the sodium phosphate buffer. | ||
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.0015 g of inosine × <math>\frac{1 mol}{268.2 g}</math> = .000005596 mol ÷ .001 L = .005596 M = 5.596 mM | .0015 g of inosine × <math>\frac{1 mol}{268.2 g}</math> = .000005596 mol ÷ .001 L = .005596 M = 5.596 mM | ||
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5.596 mM (V<sub>1</sub>)= 0.1 mM (5 mL) | 5.596 mM (V<sub>1</sub>)= 0.1 mM (5 mL) | ||
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V<sub>1</sub> = 0.08934 mL = 89.3 μL in 5 mL of buffer | V<sub>1</sub> = 0.08934 mL = 89.3 μL in 5 mL of buffer | ||
| + | * 3 mM adenosine was prepared by dissolving 0.0082 g of the solid into 10 mL sodium phosphate buffer. The stock concentration of adenosine was 3.07 mM. | ||
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| + | .0082 g of adenosine × <math>\frac{1 mol}{267.24 g}</math> = 3.06840 × 10<sup>-5</sup> ÷ .010 L = .00307 M = 3.07 mM | ||
| + | ==UV-visible scans of Reagents== | ||
| + | * Mody and Nagle condcuted the runs for the UV-visible scans of ADA, adenosine, and inosine to verify the absorbance peaks for each. | ||
| + | * From the [[AU Biomaterials Design Lab:Protocols/ADA Activity Assay|ADA Activity Assay]] protocol, it was specified to monitor the absorbance of each reagent at wavelengths 235 and 265. | ||
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Revision as of 05:00, 9 December 2012
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ADA Kinetic Assay Preparations
V1 = 0.08934 mL = 89.3 μL in 5 mL of buffer
UV-visible scans of Reagents
| |
= .0025 mol of Na2HPO4 × 
= 0.6702 g
= .000005596 mol ÷ .001 L = .005596 M = 5.596 mM
= 3.06840 × 10-5 ÷ .010 L = .00307 M = 3.07 mM
