A.

Void Volume Determination (Standard)
Vo = 5 mL
Void Volume
Fraction Equivalent Volume, mL
Absorbance
1
0.5
0.001
2
1
0
3
1.5
-0.001
4
2
-0.001
5
2.5
-0.003
6
3
0.095
7
3.5
0.094
8
4
0.098
9
4.5
0.113
10
5
0.122
11
5.5
0.099
12
6
0.095
13
6.5
0.091
14
7
0.096
15
7.5
0.093
16
8
0.092
17
8.5
0.092
18
9
0.091
19
9.5
0.094
20
10
0.093
21
10.5
0.093
22
11
0.09
23
11.5
0.096
24
12
0.092
25
12.5
0.091
26
13
0.089
27
13.5
0.09
28
14
0.09
29
14.5
0.091
30
15
0.089
31
15.5
0.09
32
16
0.089
33
16.5
0.088
34
17
0.087
35
17.5
0.092
36
18.5
0.086
37
19.5
0.099
38
20.5
0.088
39
21.5
0.085
40
22.5
0.089
41
23.5
0.083
42
24.5
0.086

B. Elution Volume Determination
Protein Standard Solution
Fraction Elution Volume, mL Absorbance
1
0.5
0
2
1
0.001
3
1.5
0.017
4
2
0.004
5
2.5
0.004
6
3
0.021
7
3.5
0.016
8
4
0.085
9
4.5
0.355
10
5
0.423
11
5.5
0.31
12
6
0.279
13
6.5
0.243
14
7
0.224
15
8
0.137
16
9
0.116
17
10
0.052
18
11
0.04
19
12
0.051
20
13
-0.061
21
14
-0.039
22
15
0.075
23
16
0.08
24
17
0.063
25
18
0.051
26
19
0.034
27
20
0.015
28
21
0.012
29
22
0.052
30
23
0.042
31
24
0.084
32
25
0.066
33
26
0.029
34
27
0.056
35
28
0.037
36
29
0.049
37
30
0.075
38
31
0.042
39
32
0.027
40
33
0.029
41
34
0.03
42
35
0.028

5 0.43 44 45 46 47 48 49 50 51 25.083 0.5 32.5 30.5 28.5 33.083 0.5 31.084 .086 0.083 0.5 26.5 27.085 0.083 0.089 0.5 29.086 0.

245 17 9 0.21 4 2 0. Elution Volume Determination Fraction Protein Extract Solution Elution Volume. Void Volume Determination (Samples) Vo = 4.5 0. mL Absorbance 1 0.5 0.5 0.2 20 10.102 2 1 0.406 12 6.173 15 8 0.07 7 3.5 0. mL 1 1 2 2 3 3 4 4 5 5 6 6 7 7 8 8 9 9 10 10 11 11 12 12 13 13 14 14 15 15 16 16 17 17 18 18 19 19 20 20 21 21 22 22 23 23 24 24 25 25 26 26 27 27 28 28 29 29 30 30 .5 0.5 0.5 0.5 0.161 3 1.507 11 6 0.5 0.536 9 5 0.5 0.139 16 8.054 8 4.519 10 5.5 mL Void Volume Fraction Elution Volume.31 14 7.165 19 10 0.5 0.153 6 3 0.233 13 7 0.259 18 9.115 5 2.15 D.C.

.

4 0.6 0.02 0 5 10 15 20 25 Equivalent Volume.54 1.02 0 -0.006 -0.mination 0.012 -0.08 0.01 -0.207 -0.12 Absorbance 0.5 Absorbance 0.5 Absorbance act Solution Absorbance 0.451 1. mL Vo Determination (Samples) 0.06 Series1 0.29 2.044 0.201 -0.161 -0.14 0. mL Ve Determination (Samples) 12 .1 0 0 10 -0.207 -0.3 Series1 0.012 -0.206 -0.119 0.281 0.219 -0.175 0.1 0 0 2 4 6 8 10 Equivalent Volume. mL 30 35 Ve Determination (Standards) 0.1 0.04 0.4 0.216 -0.178 -0.203 0.156 -0.188 -0.494 0.688 2.2 0.2 -0.158 0.22 Vo Determination (Standards) 0.1 20 30 40 Equivalent Volume.3 0.2 -0.2 Series1 0.123 0 -0.

0.5 9.5 0 5 10 15 20 Equivalent Volume. 2.5 Series1 1 18.451 Absorbance 2 1.Ve Determination (Samples) 3 2.5 0 -0. mL 25 30 35 .2 0.

093422 Calibration Curve 6 log MW 5 4 y = -0.Protein Standard Alcohol dehydrogenase Bovine serum albumin Carbonic anhydrase Cytochrome C Molecular Weight. Da 150000 66000 29000 12400 Ve 5 15 24 30 Ve/Vo 1 3 4.462398 4.176091 4.2119x + 5.9864 3 Series1 2 Linear (Series1) 1 0 0 2 4 Ve/Vo 6 8 .8 6 log MW 5.4218 R² = 0.819544 4.

Protein Extract Albumin Casein Molecular Weight. Da 45000 24000 Ve 9 18 Ve/Vo 2 4 log MW 4.31071811 .998 4.5742 MW Percent error 99540.54 121.57 56.2012039 37514.

015 0.[KCl] 0 0.1 0.5 0.5 1 0.5 Average 2.015 0.265 2.015 2.3 0.021 0.014 0.009 0.018 0.5 1 1 Absorbance Trial 1 Trial 2 2.2 0.5 0 0 -0.5 0.4 .016 0.012 0.0145 0.242 0.2 0.27 2.012 0.2 0.1985 0.002 0.193 0.01 0.009 0.0165 0.01 0.0055 0.26 2.27 0.009 0.011 2 Absorbance Fraction 1 2 3 4 5 6 7 8 9 10 11 1.3 0.1 0.214 2.204 0.007 0.016 0.013 0.

8 1 1.6 [KCl] 0.2 .Series1 0.

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