M- Protein Determination (Bradford Assay) and Photometry

M- Protein Determination (Bradford Assay) and Photometry -...

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Melody Samuel-Boutros PHS 2101- 8:00-10:50 AM Title : Protein Determination (Bradford Assay) and Photometry Objective : To use the Bradford Assay to determine the concentration of the proteins. The Bradford Assay is used in two ways- standard assay and micro assay. Methods : Refer to lab handout, page 8 Results: Test Tube # Volume PBS Volume BSA Final Conc. Of BSA Final Abs. of BSA 1 4.0 mL 0.0 mL Blank 0.0 2 3.0 mL 1.0 mL 6.25 0.273 3 2.0 mL 2.0 mL 12.5 0.436 4 1.0 mL 3.0 mL 18.75 0.590 5 0.0 mL 4.0 mL 25.0 0.730 6 Unknown 6.99 0.249 Buffer, pH Absorption KH 2 PO 4 / NaOH, pH 6.0 [unionized] 0.037 KH 2 PO 4 / NaOH, pH 7.4 [equilibrium] 0.343 Carbonate, pH 9.0 [ionized] 0.458 Carbonate, pH 10.0 [ionized] 0.486 Concentration vs. Absorbance y = 0.0284x + 0.0504 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0 5 10 15 20 25 30 Concentration Absorbance y= 0.0284x + 0.0504 0.249= 0.0284x + 0.0504 6.99= x
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Discussion and References : In the first part of this lab we saw the concentration of BSA increase. As the concentration of BSA increased, the absorbance increased. This may
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M- Protein Determination (Bradford Assay) and Photometry -...

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