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To measure the amount of protein in your unknown sample, you first need to calibrate the change in bradford reagent absorbance induced by different amount of protein.
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You already did this in lab.
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When you determine extinction, coefficiency of bsa bound to the bradford assay, in your own word, explain the theory behind how we determine the total protein concentration in our samples.
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So the very first step, you need to use the bsa standard with non -concentration to make a standard curve.
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So for example, you have five different bsa sample.
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They have different dilutions.
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So then you start with 2 mg per ml, 1 mg per ml, 0 .5 mg per ml, 0 .25, and 0.
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So these are concentrations that we know of.
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And then we use a spectrophotometer to test the measured absorbance of each one.
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So let's say you'll have absorbance.
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So these are the absorbance of each of those samples from spectrophotometer.
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So now we know that the concentration here is going to be x and absorbance is going to be y.
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So you correlate the x and y to make a standard curve.
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So absorbance is at 595 nanometer.
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Y is going to be the absorbance at 595.
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X is going to be the concentration, milligram per ml.
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So then you make a curve correlate x and y.
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So each x give you a specific y like that.
03:06
So this is concentration.
03:12
And then you make a curve according to the x and y correlation...