4. A Crude extract is diluted by taking 0.25 ml and bringing it to a total volum

Question

4. A Crude extract is diluted by taking 0.25 ml and bringing it to a total volume of 4.0 ml The following data was obtained A280 0.79 A260 0.22 Bradford http://www.csun.edu/hcchm001/Bradford.PDF A5950m Protein (ug) 0.0 1.6 3.2 4.8 6.4 8.0 9.6 5 ul of the diluted sample generated an As9s of 0.39 0.00 0.10 0.21 0.33 0.44 0.52 0.62 Determine the protein concentration of this crude extract and compare the values resulted from different calculation methods. Convert mg/mL to mM, assuming the Molecular Mass/Molecular Weight was 72,000 daltons

Explanation / Answer

The concentration of protein is calculated by the following formula,

= (1.55 × A280) - (0.76 x A260)

= (1.55 × 0.79) - (0.76 × 0.22)

= 1.2245 - 0.1672

= 1.0573 mg/ml

The crude extract was diluted to 1:16 times. So, the concentration of protein in crude extract is,

= 1.0573 × 16

= 16.917 mg/ml

= (16.917/72000) × 1000 mM

= 16917/72000 mM

= 0.235 mM

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