Rabia Iqbal
Biol 395-1005 lab
01/31/2015
Broadford assay Absorbance
Figure 1. The concentration and absorbance of Bradford assay. Four samples containing Bovine serum albumin, diluent, and Bradford. At 0.25 mg/ml concentration the absorbance was 0.109 nm and the relation between absorbance and the concentration held constant throughout except at the 0.50 mg/ml when the concentration absorbance was 0.186 nm. Figure showed that as the concentration is increased the absorbance elevates also. All the samples were in best fit line and R2=0.99591.
Calculations:
The unknown was (#8) and the absorbance was 0.105 (OD), and calculated concentration was 0.258 (mg/ml). The calculations were obtained by using the formula y=mx+b to solve for x and excel …show more content…
The dilutent was distilled water. The BSA was placed on the ice prior to being mixed into the four micro-tubes mentioned before.
The 20ml dilution was added into the four micro-centrifuge tubes including the control. Two additional tubes were added, one with 20ml of distilled water and the other was unknown. After that, 980 micro liter diluted Bradford dye was added into the four microtubes and vortexed for proper mixture, five or ten minutes’ wait was followed to ensure a better read.
The spectrophotometer was used to analyze the absorbance of protein in five micrometer tubes including the unknown after 5-10 minutes. The unknown absorbance found out from the spectrophotometer was used to figure out the concentration of the unknown protein. The wavelength was 595nm for all five tubes tubes including the unknown. Before the actual reading of the concentration in the tube, spectrophotometer was blanked (zero nanometer) by adding distilled water (control) to eliminate any reading discrepancies. The reading of concentrations was ordered based on the most dilute to the
Biol 395-1005 lab
01/31/2015
Broadford assay Absorbance
Figure 1. The concentration and absorbance of Bradford assay. Four samples containing Bovine serum albumin, diluent, and Bradford. At 0.25 mg/ml concentration the absorbance was 0.109 nm and the relation between absorbance and the concentration held constant throughout except at the 0.50 mg/ml when the concentration absorbance was 0.186 nm. Figure showed that as the concentration is increased the absorbance elevates also. All the samples were in best fit line and R2=0.99591.
Calculations:
The unknown was (#8) and the absorbance was 0.105 (OD), and calculated concentration was 0.258 (mg/ml). The calculations were obtained by using the formula y=mx+b to solve for x and excel …show more content…
The dilutent was distilled water. The BSA was placed on the ice prior to being mixed into the four micro-tubes mentioned before.
The 20ml dilution was added into the four micro-centrifuge tubes including the control. Two additional tubes were added, one with 20ml of distilled water and the other was unknown. After that, 980 micro liter diluted Bradford dye was added into the four microtubes and vortexed for proper mixture, five or ten minutes’ wait was followed to ensure a better read.
The spectrophotometer was used to analyze the absorbance of protein in five micrometer tubes including the unknown after 5-10 minutes. The unknown absorbance found out from the spectrophotometer was used to figure out the concentration of the unknown protein. The wavelength was 595nm for all five tubes tubes including the unknown. Before the actual reading of the concentration in the tube, spectrophotometer was blanked (zero nanometer) by adding distilled water (control) to eliminate any reading discrepancies. The reading of concentrations was ordered based on the most dilute to the