Tube

No.

The BSA used has a concentration of

mL BSA
Conc. of Absorbance
mL H20
protein

1
2
3
4
5
6
7
8

0
0.10
0.15
0.20
0.25
0.30
0.35
0.40

1.50
1.40
1.35
1.30
1.25
1.20
1.15
1.10

0
3.33
5
6.67
8.33
10
11.67
13.33

0.249
0.390
0.786
1.304
0.800
0.724
1.063
0.925

9
10

0.45
Unknow
n

1.05
Unknow
n

15
X

0.692
1.200

100g/mL.
The
following
concentration of the protein
samples and their absorbance:
Table
1.
absorbance

Concentration

of

are
from

protein

the
the
and

The Bradford assay is usually used to

determine the total protein concentration of
a sample. In determining the total protein
concentration of the sample, the dilution
formula was used to compute for it. The
formula is shown below:

C1 V 1=C 2 V 2

To determine the concentration of the

unknown protein, graphing method can be
used in getting the concentration. The
protein concentration is in the x-axis while
absorbance was in the y-axis.
Due to some errors in the experiment,
the most ideal line in protein standard curve
was not linear. The line was not necessarily
the best line through the origin and the other
points. With that, the unknown protein
concentration was determined by tracing the
point in which the absorbance, 1.200, met
the best fit line of the graph. The
concentration was approximately measured
to be 31.18 g /mL .
On the other hand, to get a more
accurate result, the slope and y-intercept
was determined using the linear regression
method. The equation is shown below;
wherein y is the value of absorbance, m is
the slope and b is the y-intercept.

y=mx +b

wherein,
1.5

C1 = concentration
V1 = volume of the
C2 = concentration
with water
V2 = volume of the
water

of the sample
sample
of the sample diluted
sample diluted with

1
Absorbance

f(x) = 0.01x + 0.7

R = 0.05

0.5
0
2

8 10 12 14 16

Protein Concentration g/mL

Absorbance

5 10 15 20 25 30 35

Protein Concentration

Fig 1. Albumin Standard Curve

Fig 2. Slope-intercept Equation

Using a calculator, the values of slope
and y-intercept were determined with the
use of known x, protein concentration of the
standard, and y absorbance. The value for m
is 0.014, b is 0.703 and y is 1.200

x=

x=

yb
m
1.2000.706
0.016

x=3 4.503 g /mL

Fig 3. Computation for Protein
Concentration using Linear Regression