Professional Documents
Culture Documents
2A-PH, Group 2, Ballada, T., Banares, S., Belen, R., Bergonia R., Caraig, R.Z., and Cana, N.
Department of Pharmacy, Faculty of Pharmacy, University of Santo Tomas,
Espaa Boulevard, 1015 Manila, Philippines
ABSTRACT
Identification of a substance by its protein concentration can be done
by spectrophotometry and Bradford assay, which includes combining
Coomassie G-250 dye with protein solutions. Six test tubes were prepared
with different volume of standard and
water. After subjecting to a
spectrophotometer, the absorbance of each sample was determined. The
dye used changed to darker colors when in contact with a higher protein
concentration, a higher absorbance meant a higher concentration.
Correlation between absorbance and concentration is determined, R2 is
equal to 0.9721 Also, the line of best fit was calculated, y = 0.0153x +
0.0052.
INTRODUCTION
The Bradford assay is a fast and
fairly accurate method for the estimation of
protein concentration.
METHODOLOGY
A series of test tube were prepared as
follows:
1
2
3
4
5
6
mL
0
0.2 0.4 0.6 0.8 1.0
standar
d
mL H2O 1.5 1.3 1.1 0.9 0.7 0.5
Table 1. Different volumes of standard and
voumes of H2O in different test tubes
The BSA standard was diluted to 10
mcg/mL. Then, 1.5 mL of Bradford reagent
was added to the individual test tube and
was allowed to stand for 5 minutes. The
samples were transferred into individual
cuvettes and were inserted to the
spectrophotometer. At, 595 nm, the
absorbance was read. The data was gathered.
RESULTS AND DISCUSSIONS
The results in Table 2 shows the data
gathered in Bradford assay after undergoing
the UV-vis spectrophotometer.
mL [BSA]
BSA ug/mL ABS
0.0 0.000 0.000
0.2 1.33 0.032
0.4 2.67 0.052
0.6 4.00 0.061
0.8 5.33 0.080
1.0 6.67 0.112
the
concentration
and
the
REFERENCES
Crisostomo, A. & et. al. (2010). Laboratoy
General Biochemistry. Quezon City: C & E
Publishing, Inc.