00:01
What a technician do in order to determine the protein concentration in one of the sample.
00:05
So i'm going to give you a general concept of standard curve.
00:16
So what we have is that usually to generate a standard curve, you will have certain protein standard with non -concentration.
00:24
Let's for example, you have five bovine arbumin, let's say bovine serum arbumin.
00:37
This is a very abundant protein we call this bsa sample.
00:41
Now we know these five are we called protein standard and we know that the concentration of each sample.
00:56
So you have let's say a, b, c, d and e.
01:01
And you know the concentration, let's say 2 milligram, 1, 0 .5, 0 .25 and 0 milligram per mil.
01:15
Then you are going to mix this protein sample with a reagent, we called biuret reagent.
01:32
The biuret reagent basically will turn the sample purple, and the amount of protein actually is proportional to the purple color.
01:43
So when you have a higher amount of protein, that means you have a darker purple color.
01:48
And then you measure these example by a spectrometer or a microplate reader you will have absorbance value so you will have let's say the highest here 0 .8 or 8 here and second is 0 .5 0 .2 0 .1 and obviously the last one is 0.
02:11
So then you are going to make a standard curve by use excel.
02:16
With x, the concentration, and the wide absorbance value, you are going to make a curve.
02:35
So you can see that this is 1 .8 .6 .4 .2.
02:42
This is that say, point 2, 0 .4, 0 .6 .8, 1.
03:11
Just make a curve according to concentration and absorbance value.
03:19
So then you can see that we can make a curve.
03:43
So this is a, b, c, d, and e...