Expt3 Bradford Assay- Sample Journal Article Report F09

After this 100 l of deionized water were pipeted into

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Unformatted text preview: f the previously made solutions in two new separate test tubes. After this, 100 μL of deionized water were pipeted into a test tube labeled BLANK to serve as the blank for the experiment, and 100 μL of each of the solutions of BSA of unknown concentration into separate, clean, dry, and appropriately labeled test tubes. After gently mixing and shaking the dye reagent in its bottle, 3.0 mL of Bradford reagent (St. Louis, Mo) were added to each of the test tubes. The contents of each test tube were then mixed with the dye and allowed to incubate for 5 minutes. At this point, the spectrophotometer (Fullerton, CA) was configured to read at a fixed wavelength of 595 nm, and the spectrophotometer was blanked using the BLANK tube containing Bradford Reagent and water. Following this, the absorbencies of the standard dilutions were measured and recorded, starting from the least concentrated going to the most concentrated, using the disposable cuvettes. The absorbencies of the dilutions of the BSA unknown, A1, B1, A2, and B2, were also measured and recorded. However, because the A2 readings were inconsistent with those of B2, a second sample of A2 was made using the same procedure as before, and the new readings were more in line with those of B2. Results: Attached to the report is the raw experimental data Serial Dilution Table: Sample Volume of Water (μL) 1 .5 .2 2 .4 .3 3 .4 .2 4 .3 .3 5 .3 .3 6 .3 .3 *from 2000 μg/ml BSA stock solution Sample Stock A2 B2 A1 B1 Unknown Volume of BSA (μL) Concentration Dilution of BSA Factor (μg/mL) 1428 5/7 816 4/7 544 2/3 272 ½ 136 ½ 68 ½ Dilution Factor --50x 50x 10x 10x --- Ove...
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