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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
*from 2000 μg/ml BSA stock solution
Unknown Volume of
BSA (μL) Concentration Dilution
½ Dilution Factor
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- Spring '13