Protein Lab

Protein Lab - 1 Nick Baccari Graham Barnett Jillian Daviau...

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Nick Baccari Graham Barnett Jillian Daviau Section 2 Total Protein Determination Objectives: The purpose of this experiment was to determine the amount of protein in a unknown serum complex by using known concentrations of Bovine Serum Albumen (BSA) to create a calibration curve at an absorbance scale at 595 NM. Materials: Reagents: BSA stock solution at 2 mg/mL in 20 mM Tris-HCI pH 7.5. Two unknown samples containing important serum proteins such as albumin, immunoglobulins, and various enzymes. Concentrated Bradford reagent. Buffer 10 mM Tris-HCI pH 7.5. Glassware: Large test tubes 5 Small test tubes 8 Graduated cylinders 10 mL and 100 mL Cuvettes for spectrophotometer 8 Graduated 1 mL pipet Data: Table 1: BSA-DYE STANDARDS (mg/ml BSA) Transmittance @ 595 NM Absorbance @ 595 NM Color 0 100 0 Brown "ice tea color" 0.2 60.5 0.22 Brown with slight greenish tinge 0.5 35 0.46 Dirty Blue with greenish tinge 0.7 18 0.75 Dirly Dark Blue 0.9 12 0.95 Dark Blue 1.2 9 1.03 Very Blue Unknown 1 0.5 0.98 Dark Blue (estimated between 0.7 and 0.9) Unknown 2 17 0.76 Dirty Blue (estimated between 0.5 and 0.7) Graph 1: See Attached Calculations: The slope of the protein absorbance was determined by Excel. By using the equation provided the amount of absorption for any BSA concentration of X could be determined by plugging in any protein concentration into the equation. Inversely to find
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Protein Lab - 1 Nick Baccari Graham Barnett Jillian Daviau...

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