Table 5)
influencing factors
1 staining time
2 destaining time
3 temperature of staining/staining solutions
4 temperature of selectrophoretic process
5 sample buffer
6 age of gels
7 gels
parameters
1h
2h
fridge
fridge
SDS
old
precasted
+
5h
20 h
room temperature
room temperature
LDS
new
selfcasted
mean
+
+
+
+
+
+
+
+
+
+
+
+
1
+
+
+
+
+
+

2
+
+
+
+
+
+

no.
mean
1
2
3
4
5
6
7
8
9
10
11
12
sum +
sum total sum
difference
effect
+
+
+
+
+
+
+
+
+
+
+
+
129.80
129.80
129.80
21.6333
1
staining
time
+
+
+
+
+
+
69.00
60.80
129.80
8.20
1.3667
2
destaining
time
+
+
+
+
+
+
89.80
40.00
129.80
49.80
8.3000
3
+
+
+
+
+
+

Galactosidase
influencing factors
3
temp. of
staining
+
+
+
+
+
+
49.70
80.10
129.80
30.40
5.0667
Equation 2
Ex =
x(+)
N/2
x()
N/2
2. Calculation of the experimental error using the effects of the dummy factors
2.1. the square of dummy effects were summed up:
2.2. the variance of the effects was calculated by dividing this sum by the number of dummy ef
2.3. the standard deviation of the effects was calculated by taking the square root of the varian
3. Statistical interpretation
the Minimum Significant Factor Effect (MIN) is the limit value which represents the experimenta
and was calculated by multiplying the standard deviation with a particular tvalue
tvalue:
probability value :
degrees of freedom
= number of dummy factors:
1.533
0.2
4
Phosphorylase b
no.
mean
1
2
3
4
5
6
7
8
9
10
11
12
sum +
sum total sum
+
+
+
+
+
+
+
+
+
+
+
+
115.90
115.90
1
staining
time
+
+
+
+
+
+
61.75
54.15
115.90
influencing factors
2
3
destaining temp. of
time
staining
+
+
+
+
+
+
+
+
+
+
+
+
56.60
36.30
59.30
79.60
115.90
115.90
difference
effect
115.90
19.3167
7.60
1.2667
2.70
0.4500
43.30
7.2167
Ex =
x(+)
N/2
x()
N/2
2. Calculation of the experimental error using the effects of the dummy factors
2.1. the square of dummy effects were summed up:
2.2. the variance of the effects was calculated by dividing this sum by the number of dummy ef
2.3. the standard deviation of the effects was calculated by taking the square root of the varian
3. Statistical interpretation
the Minimum Significant Factor Effect (MIN) is the limit value which represents the experimenta
and was calculated by multiplying the standard deviation with a particular tvalue
tvalue:
probability value :
degrees of freedom
= number of dummy factors:
1.533
0.2
4
BSA
no.
mean
1
2
3
4
5
6
7
8
9
10
+
+
+
+
+
+
+
+
+
+
1
staining
time
+
+
+
+
+
+
2
destaining
time
+
+
+
+
+

influencing factors
3
temp. of
staining
+
+
+
+
+
11
12
sum +
sum total sum
difference
effect
+
+
108.10
108.10
108.10
18.0167
54.10
54.00
108.10
0.10
0.0167
+
58.10
50.00
108.10
8.10
1.3500
+
43.45
64.65
108.10
21.20
3.5333
Ex =
x(+)
N/2
x()
N/2
2. Calculation of the experimental error using the effects of the dummy factors
2.1. the square of dummy effects were summed up:
2.2. the variance of the effects was calculated by dividing this sum by the number of dummy ef
2.3. the standard deviation of the effects was calculated by taking the square root of the varian
3. Statistical interpretation
the Minimum Significant Factor Effect (MIN) is the limit value which represents the experimenta
and was calculated by multiplying the standard deviation with a particular tvalue
tvalue:
probability value :
degrees of freedom
= number of dummy factors:
1.533
0.2
4
Glutamic dehydrogenase
no.
mean
1
2
3
4
5
+
+
+
+
+
1
staining
time
+
+
+
+
2
destaining
time
+
+
+
+
influencing factors
3
temp. of
staining
+
+
+

6
7
8
9
10
11
12
sum +
sum total sum
difference
effect
+
+
+
+
+
+
+
90.75
90.75
90.75
15.1250
+
+
42.25
48.50
90.75
6.25
1.0417
+
+
43.10
47.65
90.75
4.55
0.7583
+
+
+
45.45
45.30
90.75
0.15
0.0250
Ex =
x(+)
N/2
x()
N/2
2. Calculation of the experimental error using the effects of the dummy factors
2.1. the square of dummy effects were summed up:
2.2. the variance of the effects was calculated by dividing this sum by the number of dummy ef
2.3. the standard deviation of the effects was calculated by taking the square root of the varian
3. Statistical interpretation
the Minimum Significant Factor Effect (MIN) is the limit value which represents the experimenta
and was calculated by multiplying the standard deviation with a particular tvalue
tvalue:
probability value :
degrees of freedom
= number of dummy factors:
1.533
0.2
4
Lactate dehydrogenase
no.
mean
1
staining
time
2
destaining
time
influencing factors
3
temp. of
staining
1
2
3
4
5
6
7
8
9
10
11
12
sum +
sum total sum
difference
effect
+
+
+
+
+
+
+
+
+
+
+
+
59.20
59.20
59.20
9.8667
+
+
+
+
+
+
31.05
28.15
59.20
2.90
0.4833
+
+
+
+
+
+
29.45
29.75
59.20
0.30
0.0500
+
+
+
+
+
+
29.45
29.75
59.20
0.30
0.0500
Ex =
x(+)
N/2
x()
N/2
2. Calculation of the experimental error using the effects of the dummy factors
2.1. the square of dummy effects were summed up:
2.2. the variance of the effects was calculated by dividing this sum by the number of dummy ef
2.3. the standard deviation of the effects was calculated by taking the square root of the varian
3. Statistical interpretation
the Minimum Significant Factor Effect (MIN) is the limit value which represents the experimenta
and was calculated by multiplying the standard deviation with a particular tvalue
tvalue:
probability value :
degrees of freedom
= number of dummy factors:
1.533
0.2
4
Carbonic anhydrase
no.
mean
1
2
3
4
5
6
7
8
9
10
11
12
sum +
sum total sum
difference
effect
+
+
+
+
+
+
+
+
+
+
+
+
62.25
62.25
62.25
10.3750
1
staining
time
+
+
+
+
+
+
34.00
28.25
62.25
5.75
0.9583
2
destaining
time
+
+
+
+
+
+
34.35
27.90
62.25
6.45
1.0750
influencing factors
3
temp. of
staining
+
+
+
+
+
+
27.65
34.60
62.25
6.95
1.1583
Ex =
x(+)
N/2
x()
N/2
2. Calculation of the experimental error using the effects of the dummy factors
2.1. the square of dummy effects were summed up:
2.2. the variance of the effects was calculated by dividing this sum by the number of dummy ef
2.3. the standard deviation of the effects was calculated by taking the square root of the varian
3. Statistical interpretation
the Minimum Significant Factor Effect (MIN) is the limit value which represents the experimenta
and was calculated by multiplying the standard deviation with a particular tvalue
tvalue:
probability value :
degrees of freedom
1.533
0.2
Trypsin inhibitor
no.
mean
1
2
3
4
5
6
7
8
9
10
11
12
sum +
sum total sum
difference
effect
+
+
+
+
+
+
+
+
+
+
+
+
80.95
80.95
80.95
13.4917
1
staining
time
+
+
+
+
+
+
39.65
41.30
80.95
1.65
0.2750
influencing factors
2
3
destaining temp. of
time
staining
+
+
+
+
+
+
+
+
+
+
+
+
39.45
41.95
41.50
39.00
80.95
80.95
2.05
2.95
0.3417
0.4917
Ex =
x(+)
N/2
x()
N/2
2. Calculation of the experimental error using the effects of the dummy factors
2.1. the square of dummy effects were summed up:
2.2. the variance of the effects was calculated by dividing this sum by the number of dummy ef
2.3. the standard deviation of the effects was calculated by taking the square root of the varian
3. Statistical interpretation
the Minimum Significant Factor Effect (MIN) is the limit value which represents the experimenta
and was calculated by multiplying the standard deviation with a particular tvalue
tvalue:
probability value :
degrees of freedom
= number of dummy factors:
1.533
0.2
4
Two further proteins (Myosin and Lysozym) were investigated. However, these proteins appear
areas was not possible in these cases. Therefore, these proteins were not used for the evaluat
For completeness the results of these proteins are shown in the following two tables:
Myosin
no.
mean
1
2
3
4
5
6
7
8
9
10
11
12
sum +
sum total sum
difference
sum + / n +
sum  / n effect
+
+
+
+
+
+
+
+
+
+
+
+
128.55
128.55
128.55
21.4250
1
staining
time
+
+
+
+
+
+
62.80
65.75
128.55
2.95
15.70
13.15
2.5500
influencing factors
2
3
destaining temp. of
time
staining
+
+
+
+
+
+
+
+
+
+
+
+
68.30
59.50
60.25
69.05
128.55
128.55
8.05
9.55
17.08
11.90
12.05
17.26
5.0250
5.3625
Ex =
x(+)
N/2
x()
N/2
2. Calculation of the experimental error using the effects of the dummy factors
2.2. the variance of the effects was calculated by dividing this sum by the number of dummy ef
2.3. the standard deviation of the effects was calculated by taking the square root of the varian
3. Statistical interpretation
the Minimum Significant Factor Effect (MIN) is the limit value which represents the experimenta
and was calculated by multiplying the standard deviation with a particular tvalue
tvalue:
probability value :
degrees of freedom
= number of dummy factors:
Lysozym
no.
1
2
3
4
5
6
7
8
9
10
11
12
sum +
sum total sum
difference
sum + / n +
sum  / n effect
mean
+
+
+
+
+
+
+
+
+
+
+
+
60.2
60.2
60.2
10.0333
1
staining
time
+
+
+
+
+
+
39.3
21.0
60.2
18.3
7.9
4.2
3.6600
1.533
0.2
4
2
destaining
time
+
+
+
+
+
+
20.6
39.7
60.2
19.1
5.1
6.6
1.4708
Ex =
x(+)
N/2
x()
N/2
influencing factors
3
temp. of
staining
+
+
+
+
+
+
29.9
30.4
60.2
0.5
6.0
6.1
0.1000
Ex =
x(+)
N/2
x()
N/2
2. Calculation of the experimental error using the effects of the dummy factors
2.1. the square of dummy effects were summed up:
2.2. the variance of the effects was calculated by dividing this sum by the number of dummy ef
2.3. the standard deviation of the effects was calculated by taking the square root of the varian
3. Statistical interpretation
the Minimum Significant Factor Effect (MIN) is the limit value which represents the experimenta
and was calculated by multiplying the standard deviation with a particular tvalue
tvalue:
probability value :
degrees of freedom
= number of dummy factors:
1.533
0.2
4
4
+
+
+
+
+
+

influencing factors
4
temp. of
electroph.
+
+
+
+
+
+
70.45
59.35
129.80
11.10
1.8500
5
+
+
+
+
+
+

6
+
+
+
+
+
+

7
+
+
+
+
+
+

8
+
+
+
+
+
+

5
sample
buffer
+
+
+
+
+
+
72.15
57.65
129.80
14.50
2.4167
6
age of
gels
+
+
+
+
+
+
65.15
64.65
129.80
0.50
0.0833
7
gels
+
+
+
+
+
+
62.65
67.15
129.80
4.50
0.7500
+
+
+
+
+
+
70.65
59.15
129.80
11.50
1.9167
9
+
+
+
+
+
+

10
+
+
+
+
+
+

11
+
+
+
+
+
+

dummy factors
9
10
11
+
+
+
+
+
+
71.40
58.40
129.80
13.00
2.1667
+
+
+
+
+
+
59.15
70.65
129.80
11.50
1.9167
+
+
+
+
+
+
54.30
75.50
129.80
21.20
3.5333
The effect was calculated by determining the difference between the average
of the + and  responses Y for each independent and dummy variable.
N is the number of experiments of the design (in this case N = 12 experiments
consisting of 6 + and  experiments, respectively)
sum:
3.6736
4.6944
3.6736
12.4844
24.5261
6.1315
2.4762
influencing factors
4
temp. of
electroph.
+
+
+
+
+
+
50.75
65.15
115.90
equation 4
MIN =
MIN:
3.7965
MIN = Minimun Significant Factor Effect
5
sample
buffer
+
+
+
+
+
+
74.60
41.30
115.90
6
age of
gels
+
+
+
+
+
+
48.40
67.50
115.90
7
gels
+
+
+
+
+
+
71.05
44.85
115.90
+
+
+
+
+
+
77.65
38.25
115.90
dummy factors
9
10
+
+
+
+
+
+
55.45
60.45
115.90
+
+
+
+
+
+
57.40
58.50
115.90
11
+
+
+
+
+
+
62.25
53.65
115.90
14.40
2.4000
33.30
5.5500
19.10
3.1833
26.20
4.3667
39.40
6.5667
5.00
0.8333
1.10
0.1833
8.60
1.4333
The effect was calculated by determining the difference between the average
of the + and  responses Y for each independent and dummy variable.
N is the number of experiments of the design (in this case N = 12 experiments
consisting of 6 + and  experiments, respectively)
sum:
43.1211
0.6944
0.0336
2.0544
45.9036
11.4759
3.3876
influencing factors
4
temp. of
electroph.
+
+
+
+
+
+
equation 4
MIN =
MIN:
5.1939
MIN = Minimun Significant Factor Effect
5
sample
buffer
+
+
+
+
+

6
age of
gels
+
+
+
+
+
7
gels
+
+
+
+
+
+
+
+
+
+
+
dummy factors
9
10
+
+
+
+
+
+

+
+
+
+
+
+

11
+
+
+
+
+

49.10
59.00
108.10
9.90
1.6500
+
52.70
55.40
108.10
2.70
0.4500
+
58.80
49.30
108.10
9.50
1.5833
+
45.05
63.05
108.10
18.00
3.0000
66.60
41.50
108.10
25.10
4.1833
56.95
51.15
108.10
5.80
0.9667
44.90
63.20
108.10
18.30
3.0500
+
51.65
56.45
108.10
4.80
0.8000
The effect was calculated by determining the difference between the average
of the + and  responses Y for each independent and dummy variable.
N is the number of experiments of the design (in this case N = 12 experiments
consisting of 6 + and  experiments, respectively)
sum:
17.5003
0.9344
9.3025
0.6400
28.3772
7.0943
2.6635
influencing factors
4
temp. of
electroph.
+
+
+

equation 4
MIN =
MIN:
4.0837
MIN = Minimun Significant Factor Effect
5
sample
buffer
+
+

6
age of
gels
+
+
7
gels
+

+
+
dummy factors
9
10
+
+
+
+
+
+

11
+
+
+
+
+
+
40.10
50.65
90.75
10.55
1.7583
+
+
+
+
46.95
43.80
90.75
3.15
0.5250
+
+
+
+
49.05
41.70
90.75
7.35
1.2250
+
+
+
+
+
45.45
45.30
90.75
0.15
0.0250
+
+
+
+
54.90
35.85
90.75
19.05
3.1750
+
+
+
50.45
40.30
90.75
10.15
1.6917
+
+
+
47.80
42.95
90.75
4.85
0.8083
+
+
+
47.20
43.55
90.75
3.65
0.6083
The effect was calculated by determining the difference between the average
of the + and  responses Y for each independent and dummy variable.
N is the number of experiments of the design (in this case N = 12 experiments
consisting of 6 + and  experiments, respectively)
sum:
10.0806
2.8617
0.6534
0.3701
13.9658
3.4915
1.8685
influencing factors
4
temp. of
electroph.
equation 4
MIN =
MIN:
2.8649
MIN = Minimun Significant Factor Effect
5
sample
buffer
6
age of
gels
7
gels
dummy factors
9
10
11
+
+
+
+
+
+
29.40
29.80
59.20
0.40
0.0667
+
+
+
+
+
+
26.70
32.50
59.20
5.80
0.9667
+
+
+
+
+
+
31.75
27.45
59.20
4.30
0.7167
+
+
+
+
+
+
27.85
31.35
59.20
3.50
0.5833
+
+
+
+
+
+
34.55
24.65
59.20
9.90
1.6500
+
+
+
+
+
+
30.85
28.35
59.20
2.50
0.4167
+
+
+
+
+
+
29.85
29.35
59.20
0.50
0.0833
+
+
+
+
+
+
33.30
25.90
59.20
7.40
1.2333
The effect was calculated by determining the difference between the average
of the + and  responses Y for each independent and dummy variable.
N is the number of experiments of the design (in this case N = 12 experiments
consisting of 6 + and  experiments, respectively)
sum:
2.7225
0.1736
0.0069
1.5211
4.4242
1.1060
1.0517
MIN:
1.6125
MIN = Minimun Significant Factor Effect
influencing factors
4
temp. of
electroph.
+
+
+
+
+
+
33.65
28.60
62.25
5.05
0.8417
5
sample
buffer
+
+
+
+
+
+
30.75
31.50
62.25
0.75
0.1250
6
age of
gels
+
+
+
+
+
+
32.50
29.75
62.25
2.75
0.4583
7
gels
+
+
+
+
+
+
30.60
31.65
62.25
1.05
0.1750
+
+
+
+
+
+
37.50
24.75
62.25
12.75
2.1250
dummy factors
9
10
+
+
+
+
+
+
30.10
32.15
62.25
2.05
0.3417
+
+
+
+
+
+
28.70
33.55
62.25
4.85
0.8083
11
+
+
+
+
+
+
31.50
30.75
62.25
0.75
0.1250
The effect was calculated by determining the difference between the average
of the + and  responses Y for each independent and dummy variable.
N is the number of experiments of the design (in this case N = 12 experiments
consisting of 6 + and  experiments, respectively)
sum:
4.5156
0.1167
0.6534
0.0156
5.3014
1.3253
1.1512
MIN:
1.7651
influencing factors
4
temp. of
electroph.
+
+
+
+
+
+
36.40
44.55
80.95
8.15
1.3583
5
sample
buffer
+
+
+
+
+
+
42.25
38.70
80.95
3.55
0.5917
6
age of
gels
+
+
+
+
+
+
48.85
32.10
80.95
16.75
2.7917
7
gels
+
+
+
+
+
+
39.95
41.00
80.95
1.05
0.1750
+
+
+
+
+
+
45.05
35.90
80.95
9.15
1.5250
dummy factors
9
10
+
+
+
+
+
+
38.70
42.25
80.95
3.55
0.5917
+
+
+
+
+
+
34.75
46.20
80.95
11.45
1.9083
11
+
+
+
+
+
+
42.70
38.25
80.95
4.45
0.7417
The effect was calculated by determining the difference between the average
of the + and  responses Y for each independent and dummy variable.
N is the number of experiments of the design (in this case N = 12 experiments
consisting of 6 + and  experiments, respectively)
sum:
2.3256
0.3501
3.6417
0.5501
6.8675
1.7169
1.3103
equation 4
MIN =
MIN:
2.0090
MIN = Minimun Significant Factor Effect
ed. However, these proteins appear at the top or rather at the bottom of the gel and a correct integration of the peak
oteins were not used for the evaluation of the PlackettBurman design.
n the following two tables:
influencing factors
4
temp. of
electroph.
+
+
+
+
+
+
51.15
77.40
128.55
26.25
10.23
19.35
9.1200
5
sample
buffer
+
+
+
+
+
+
56.20
72.35
128.55
16.15
14.05
14.47
0.4200
6
age of
gels
+
+
+
+
+
+
93.35
35.20
128.55
58.15
15.56
11.73
3.8250
7
gels
+
+
+
+
+
+
32.05
96.50
128.55
64.45
10.68
16.08
5.4000
+
+
+
+
+
+
64.60
63.95
128.55
0.65
16.15
12.79
3.3600
dummy factors
9
10
+
+
+
+
+
+
60.85
67.70
128.55
6.85
15.21
13.54
1.6725
+
+
+
+
+
+
49.65
78.90
128.55
29.25
12.41
15.78
3.3675
The effect was calculated by determining the difference between the average
of the + and  responses Y for each independent and dummy variable.
N is the number of experiments of the design (in this case N = 12 experiments
consisting of 6 + and  experiments, respectively)
11
+
+
+
+
+
+
67.85
60.70
128.55
7.15
13.57
15.18
1.6050
11.2896
2.7973
11.3401
2.5760
28.0029
7.0007
2.6459
sum:
MIN:
4.0567
MIN = Minimun Significant Factor Effect
influencing factors
4
temp. of
electroph.
+
+
+
+
+
+
37.4
22.8
60.2
14.6
7.5
4.6
2.9200
5
sample
buffer
+
+
+
+
+
+
36.6
23.7
60.2
12.9
6.1
5.9
0.1792
6
age of
gels
+
+
+
+
+
+
27.2
33.0
60.2
5.8
5.4
6.6
1.1600
7
gels
+
+
+
+
+
+
30.2
30.1
60.2
0.1
5.0
7.5
2.4875
+
+
+
+
+
+
22.6
37.6
60.2
15
5.7
6.3
0.6167
dummy factors
9
10
+
+
+
+
+
+
25.6
34.7
60.2
9.1
5.1
6.9
1.8200
The effect was calculated by determining the difference between the average
+
+
+
+
+
+
24.8
35.5
60.2
10.7
5.0
7.1
2.1400
11
+
+
+
+
+
+
23.6
36.7
60.2
13.1
5.9
6.1
0.2208
sum:
0.3803
3.3124
4.5796
0.0488
8.3210
2.0803
1.4423
MIN:
2.2114
MIN = Minimun Significant Factor Effect
response
response
RSD%
19.5
6.6
9.0
11.5
17.7
6.9
5.3
7.0
23.3
6.8
8.9
7.4
2
(Di)
Ex
> MIN
response
RSD%
7.8
5.0
5.0
4.2
12.0
26.2
6.4
7.9
20.0
6.6
7.7
7.2
2
(Di)
Ex
> MIN
response
RSD%
9.0
5.8
7.7
4.0
18.5
8.1
5.3
10.9
12.6
8.8
6.4
11.2
2
(Di)
Ex
> MIN
response
RSD%
4.3
5.4
7.4
6.1
10.2
9.4
7.9
12.8
8.2
7.0
7.0
5.3
2
(Di)
Ex
> MIN
response
RSD%
3.7
4.6
6.3
3.8
8.0
5.4
5.3
5.4
3.9
5.7
3.8
3.5
2
(Di)
Ex
> MIN
response
RSD%
6.1
4.0
5.9
3.3
8.1
5.8
3.8
3.9
7.1
6.8
3.9
3.7
2
(Di)
Ex
> MIN
Ex
> MIN
response
RSD%
5.5
5.6
5.2
3.2
9.9
6.5
5.5
8.9
5.7
9.0
10.1
6.0
2
(Di)
Ex
> MIN
on of the peak
response
RSD%
16.1
8.4
9.3
N.N.
28.2
N.N.
7.3
17.0
N.N.
10.1
14.7
17.5
2
(Di)
Ex
> MIN
response
RSD%
9.0
11.2
N.N.
3.8
N.N.
6.1
3.3
2.6
4.7
9.3
3.1
7.3
2
(Di)
Ex
> MIN