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Protein determination by the Lowry method.

Protein calibration curve.


Volume of BSA solution
0
0.1
0.2
0.2
0.3
0.5
0.5
0.8

Milligrams of protein
0
0.04
0.08
0.08
0.12
0.2
0.2
0.32

Absorbance at 750nm
0
0.094
0.154
0.199
0.234
0.421
0.455
0.550

Samples of the diluted fractions.


Sample tubes

Volume of water

1
2
3
4
5
6
7
8
9
10
11

0.99
0.98
0.95
0.99
0.98
0.95
0.99
0.98
0.95
0.7
0.5

Volume of diluted
fraction
0.01
0.02
0.05
0.01
0.02
0.05
0.01
0.02
0.05
0.3
0.5

Absorbance at 750nm
0.116
0.259
0.703
0.029
0.106
0.247
0.032
0.044
0.424
0.109
0.137

B. Assay of fractions for Invertase activity using Nelsons Procedure for reducing sugars.
Calibration curve for reducing sugars.
Glucose samples
0
0.05
0.05
0.1
0.15
0.2
0.2
0.25
0.3
0.2 (fructose)
0.2 (sucrose)

Micromoles of reducing
sugars
0
0.2
0.2
0.4
0.6
0.8
0.8
1
1.2
0.8
0.8

Absorbance at 510nm
0
0.118
0.082
0.253
0.708
0.799
0.402
0.656
0.910
0.667
0.028

Assay of fractions for Invertase activity.


Tubes
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
Graphs.

Absorbance at 510nm
0
0.560
0.591
0.500
0.350
0.525
0.592
0.057
0.225
0.312
0.060
0.136
0.125
-0.071/0
0.523

Title: Protein Calibration curve- Absorbance vs


Milligrams of protein.
1
0.9

Absorbance at 750nm.

0.8
0.7

y = 2.1076x

0.6
0.5

Series1

0.4

Linear (Series1)

0.3
0.2
0.1
0
0

0.05

0.1

0.15

0.2

0.25

Millgrams of preotein. (mg)

0.3

0.35

Title: Calibration curve of reducing sugarsAbsorbance vs Micromoles of reducing sugars.


1
0.91

0.9
y = 0.6841x
0.799

Absorbance at 510nm.

0.8
0.708

0.7

0.667

0.656

0.6
0.5

Series1

0.4

0.402

Linear (Series1)

0.3
0.253
0.2
0.118
0.082

0.1
0

0.028

0
0

0.2

0.4

0.6

0.8

Micromoles of reducing sugars. (mol)

1.2

1.4