Supporting Material_Plakett-Burman Design (1)

8/9/2019 Supporting Material_Plakett-Burman Design (1)

1/44

Investigated influencing factors and the corresponding levels (s. Table 5)

parameters

influencing factors - +

1 staining time 1 h 5 h

2 destaining time 2 h 20 h

3 temperature of staining/staining solutions fridge room temperature

4 temperature of selectrophoretic process fridge room temperature

5 sample buffer SDS LDS

6 age of gels old new

gels pre!casted self!casted


2/44


3/44


4/44


5/44


6/44


7/44


8/44


9/44


10/44


11/44


12/44

general Plackett-Burman schedule for ! e"periments (s. Table #)

no. mean ! $

" " " !

! " " ! "

$ " ! " "

# " " " "

5 " " " !

% " " ! !

& " ! ! !

' " ! ! "

" ! " !

" " ! "

" ! " "

! " ! ! !

sum "

sum !

total sum

difference

effect

*-alactosidase

influencingno. mean ! $

staining destaining temp. of

time time staining

" " " !

! " " ! "

$ " ! " "

# " " " "

5 " " " !

% " " ! !

& " ! ! !

' " ! ! " " ! " !

" " ! "

" ! " "

! " ! ! !

sum " 12#$%0 6#$00 %#$%0 4#$0

sum ! !!!!! 60$%0 40$00 %0$10

total sum 12#$%0 12#$%0 12#$%0 12#$%0

difference 12#$%0 %$20 4#$%0 !30$40

effect 21$6333 1$366 '.$ -5.%%&

. ,alculation of the effects


13/44

uation !

!. ,alculation of the e"perimental error using the effects of the du

2$1$ the s&uare of dumm' effects were summed up(

2$2$ the )ariance of the effects was calculated b' di)iding this sum b' th

2$3$ the standard de)iation of the effects was calculated b' ta*ing the s

$. 0tatistical interpretation

the +inimum Significant ,actor -ffect .+ is the limit )alue which repr

and was calculated b' multipl'ing the standard de)iation with a particul

t!)alue( 1$533

probabilit' )alue ( 0$2

degrees of freedom

number of dumm' factors( 4

Phosphor/lase b

influencing

no. mean ! $


time time staining

" " " !

! " " ! "

$ " ! " "# " " " "

5 " " " !

% " " ! !

& " ! ! !

' " ! ! "

" ! " !

" " ! "

" ! " "

! " ! ! !

sum " 115$#0 61$5 56$60 36$30

sum ! !!!!! 54$15 5#$30 #$60total sum 115$#0 115$#0 115$#0 115$#0

."1 .!1

45 5

-0/2 0/2


14/44

difference 115$#0 $60 !2$0 !43$30

effect 1#$316 1$266 !0$4500 -&.!%&


uation !








t!)alue( 1$533


degrees of freedom


B01

influencing

no. mean ! $staining destaining temp. of

time time staining

" " " !

! " " ! "

$ " ! " "

# " " " "

5 " " " !

% " " ! !

& " ! ! !

' " ! ! "

" ! " ! " " ! "

."1 .!1

45 5

-0/2 0/2


15/44

" ! " "

! " ! ! !

sum " 10%$10 54$10 5%$10 43$45

sum ! !!!!! 54$00 50$00 64$65

total sum 10%$10 10%$10 10%$10 10%$10

difference 10%$10 0$10 %$10 !21$20effect 1%$016 0$016 1$3500 !3$5333


uation !





$. 0tatistical interpretationthe +inimum Significant ,actor -ffect .+ is the limit )alue which repr


t!)alue( 1$533


degrees of freedom


lutamic deh/drogenase

influencing

no. mean ! $


time time staining

" " " !

! " " ! "

$ " ! " "

# " " " "5 " " " !

."1 .!1

45 5

-0/2 0/2


16/44

% " " ! !

& " ! ! !

' " ! ! "

" ! " !

" " ! "

" ! " "

! " ! ! !

sum " #0$5 42$25 43$10 45$45

sum ! !!!!! 4%$50 4$65 45$30

total sum #0$5 #0$5 #0$5 #0$5

difference #0$5 !6$25 !4$55 0$15

effect 15$1250 !1$041 !0$5%3 0$0250


uation !








t!)alue( 1$533probabilit' )alue ( 0$2

degrees of freedom


2actate deh/drogenase

influencing

no. mean ! $

staining destaining temp. oftime time staining

."1 .!1

45 5

-0/2 0/2


17/44

" " " !

! " " ! "

$ " ! " "

# " " " "

5 " " " !

% " " ! !

& " ! ! !

' " ! ! "

" ! " !

" " ! "

" ! " "

! " ! ! !

sum " 5#$20 31$05 2#$45 2#$45

sum ! !!!!! 2%$15 2#$5 2#$5

total sum 5#$20 5#$20 5#$20 5#$20

difference 5#$20 2$#0 !0$30 !0$30

effect #$%66 0$4%33 !0$0500 !0$0500


uation !






the +inimum Significant ,actor -ffect .+ is the limit )alue which reprand was calculated b' multipl'ing the standard de)iation with a particul

t!)alue( 1$533


degrees of freedom


."1 .!1

45 5

-0/2 0/2


18/44

,arbonic anh/drase

influencing

no. mean ! $


time time staining

" " " !! " " ! "

$ " ! " "

# " " " "

5 " " " !

% " " ! !

& " ! ! !

' " ! ! "

" ! " !

" " ! "

" ! " "

! " ! ! !

sum " 62$25 34$00 34$35 2$65

sum ! !!!!! 2%$25 2$#0 34$60

total sum 62$25 62$25 62$25 62$25

difference 62$25 5$5 6$45 !6$#5

effect 10$350 0$#5%3 1$050 !1$15%3


uation !



2$2$ the )ariance of the effects was calculated b' di)iding this sum b' th2$3$ the standard de)iation of the effects was calculated b' ta*ing the s




t!)alue( 1$533

probabilit' )alue ( 0$2degrees of freedom

."1 .!1

4

5 5

- 0/2 0/2


19/44


Tr/psin inhibitor

influencing

no. mean ! $


time time staining

" " " !

! " " ! "

$ " ! " "

# " " " "

5 " " " !

% " " ! !

& " ! ! !

' " ! ! "

" ! " !

" " ! "

" ! " "

! " ! ! !

sum " %0$#5 3#$65 3#$45 41$#5

sum ! !!!!! 41$30 41$50 3#$00

total sum %0$#5 %0$#5 %0$#5 %0$#5

difference %0$#5 !1$65 !2$05 2$#5

effect 13$4#1 !0$250 !0$341 0$4#1


uation !








."1 .!1

45 5

-0/2 0/2


20/44

t!)alue( 1$533


degrees of freedom


wo further proteins .+'osin and L'so7'm were in)estigated$ 8owe)er

areas was not possible in these cases$ herefore9 these proteins were

,or completeness the results of these proteins are shown in the followi

3/osin influencing

no. mean ! $


time time staining

" " " !

! " " ! "

$ " ! " "

# " " " "

5 " " " !

% " " ! !

& " ! ! !

' " ! ! " " ! " !

" " ! "

" ! " "

! " ! ! !

sum " 12%$55 62$%0 6%$30 5#$50

sum ! !!!!! 65$5 60$25 6#$05

total sum 12%$55 12%$55 12%$55 12%$55

difference 12%$55 !2$#5 %$05 !#$55

sum " / n " 15$0 1$0% 11$#0

sum ! / n ! 13$15 12$05 1$26

effect 21$4250 2$5500 5.!5 -5.$%!5


uation !


."1 .!1

45 5

-0/2 0/2


21/44







t!)alue( 1$533


degrees of freedom


2/so4/m influencing

no. mean ! $


time time staining

" " " !! " " ! "

$ " ! " "

# " " " "

5 " " " !

% " " ! !

& " ! ! !

' " ! ! "

" ! " !

" " ! "

" ! " "

! " ! ! !sum " 60$2 3#$3 20$6 2#$#

sum ! !!!!! 21$0 3#$ 30$4

total sum 60$2 60$2 60$2 60$2

difference 60$2 1%$3 !1#$1 !0$5

sum " / n " $# 5$1 6$0

sum ! / n ! 4$2 6$6 6$1

effect 10$0333 $.%% !1$40% !0$1000


uation !


22/44








t!)alue( 1$533


degrees of freedom


."1 .!1

4-0/2 0/2


23/44

# 5 % & '

" " " ! ! ! " !

" " ! ! ! " ! "

" ! ! ! " ! " "

! ! ! " ! " " !

! ! " ! " " ! "

! " ! " " ! " "

" ! " " ! " " "

! " " ! " " " !

" " ! " " " ! !

" ! " " " ! ! !

! " " " ! ! ! "

! ! ! ! ! ! ! !

factors dumm/ factors# 5 % & '

temp. of sample age of gels

electroph. buffer gels

" " " ! ! ! " !

" " ! ! ! " ! "

" ! ! ! " ! " "

! ! ! " ! " " !

! ! " ! " " ! "

! " ! " " ! " "

" ! " " ! " " "

! " " ! " " " !" " ! " " " ! !

" ! " " " ! ! !

! " " " ! ! ! "

! ! ! ! ! ! ! !

0$45 2$15 65$15 62$65 0$65 1$40 5#$15 54$30

5#$35 5$65 64$65 6$15 5#$15 5%$40 0$65 5$50

12#$%0 12#$%0 12#$%0 12#$%0 12#$%0 12#$%0 12#$%0 12#$%0

11$10 14$50 0$50 !4$50 11$50 13$00 !11$50 !21$20

1$%500 2$416 0$0%33 !0$500 1$#16 2$166 !1$#16 !3$5333


24/44

he effect was calculated b' determining the difference between the a)erage

of the " and ! responses for each independent and dumm' )ariable$

is the number of eperiments of the design .in this case 12 eperiments

consisting of 6 " and ! eperiments9 respecti)el'

m/ factors

3$636 he steps 2$

4$6#44

3$636 euation $

12$4%44

sum 24$5261

e number of dumm' effects 6$1315

uare root of the )ariance 2$462

esents the eperimental )ariabilit' within the design

r t!)alue

euation #

an effect is

3I6 $.&%5

+ +inimun Significant ,actor -ffect

factors dumm/ factors

# 5 % & '



" " " ! ! ! " !

" " ! ! ! " ! "

" ! ! ! " ! " "! ! ! " ! " " !

! ! " ! " " ! "

! " ! " " ! " "

" ! " " ! " " "

! " " ! " " " !

" " ! " " " ! !

" ! " " " ! ! !

! " " " ! ! ! "

! ! ! ! ! ! ! !

50$5 4$60 4%$40 1$05 $65 55$45 5$40 62$25

65$15 41$30 6$50 44$%5 3%$25 60$45 5%$50 53$65115$#0 115$#0 115$#0 115$#0 115$#0 115$#0 115$#0 115$#0

3I6 7 t


25/44

!14$40 33$30 !1#$10 26$20 3#$40 !5$00 !1$10 %$60

!2$4000 5.55 !3$1%33 4$366 6$566 !0$%333 !0$1%33 1$4333





m/ factors

43$1211 he steps 2$

0$6#44

0$0336 euation $

2$0544

sum 45$#036

e number of dumm' effects 11$45#

uare root of the )ariance 3$3%6


r t!)alue

euation #

an effect is

3I6 5.$



# 5 % & ' temp. of sample age of gels


" " " ! ! ! " !

" " ! ! ! " ! "

" ! ! ! " ! " "

! ! ! " ! " " !

! ! " ! " " ! "

! " ! " " ! " "

" ! " " ! " " "

! " " ! " " " !

" " ! " " " ! !" ! " " " ! ! !

3I6 7 t


26/44

! " " " ! ! ! "

! ! ! ! ! ! ! !

4#$10 52$0 5%$%0 45$05 66$60 56$#5 44$#0 51$65

5#$00 55$40 4#$30 63$05 41$50 51$15 63$20 56$45

10%$10 10%$10 10%$10 10%$10 10%$10 10%$10 10%$10 10%$10

!#$#0 !2$0 #$50 !1%$00 25$10 5$%0 !1%$30 !4$%0!1$6500 !0$4500 1$5%33 !3$0000 4$1%33 0$#66 !3$0500 !0$%000





m/ factors

1$5003 he steps 2$

0$#344

#$3025 euation $

0$6400

sum 2%$32

e number of dumm' effects $0#43



r t!)alue

euation #

an effect is

3I6 #.'$&



# 5 % & '



" " " ! ! ! " !

" " ! ! ! " ! "

" ! ! ! " ! " "

! ! ! " ! " " !! ! " ! " " ! "

3I6 7 t


27/44

! " ! " " ! " "

" ! " " ! " " "

! " " ! " " " !

" " ! " " " ! !

" ! " " " ! ! !

! " " " ! ! ! "

! ! ! ! ! ! ! !

40$10 46$#5 4#$05 45$45 54$#0 50$45 4$%0 4$20

50$65 43$%0 41$0 45$30 35$%5 40$30 42$#5 43$55

#0$5 #0$5 #0$5 #0$5 #0$5 #0$5 #0$5 #0$5

!10$55 3$15 $35 0$15 1#$05 10$15 4$%5 3$65

!1$5%3 0$5250 1$2250 0$0250 3$150 1$6#1 0$%0%3 0$60%3





m/ factors

10$0%06 he steps 2$

2$%61

0$6534 euation $

0$301

sum 13$#65%e number of dumm' effects 3$4#15

uare root of the )ariance 1$%6%5


r t!)alue

euation #

an effect is

3I6 !.'%#



# 5 % & '

temp. of sample age of gelselectroph. buffer gels

3I6 7 t


28/44

" " " ! ! ! " !

" " ! ! ! " ! "

" ! ! ! " ! " "

! ! ! " ! " " !

! ! " ! " " ! "

! " ! " " ! " "

" ! " " ! " " "

! " " ! " " " !

" " ! " " " ! !

" ! " " " ! ! !

! " " " ! ! ! "

! ! ! ! ! ! ! !

2#$40 26$0 31$5 2$%5 34$55 30$%5 2#$%5 33$30

2#$%0 32$50 2$45 31$35 24$65 2%$35 2#$35 25$#0

5#$20 5#$20 5#$20 5#$20 5#$20 5#$20 5#$20 5#$20

!0$40 !5$%0 4$30 !3$50 #$#0 2$50 0$50 $40

!0$066 !0$#66 0$16 !0$5%33 1$6500 0$416 0$0%33 1$2333





m/ factors

2$225 he steps 2$

0$136

0$006# euation $

1$5211

sum 4$4242

e number of dumm' effects 1$1060


esents the eperimental )ariabilit' within the designr t!)alue

euation #

an effect is

3I6 .%!5


3I6 7 t


29/44


# 5 % & '



" " " ! ! ! " !" " ! ! ! " ! "

" ! ! ! " ! " "

! ! ! " ! " " !

! ! " ! " " ! "

! " ! " " ! " "

" ! " " ! " " "

! " " ! " " " !

" " ! " " " ! !

" ! " " " ! ! !

! " " " ! ! ! "

! ! ! ! ! ! ! !

33$65 30$5 32$50 30$60 3$50 30$10 2%$0 31$50

2%$60 31$50 2#$5 31$65 24$5 32$15 33$55 30$5

62$25 62$25 62$25 62$25 62$25 62$25 62$25 62$25

5$05 !0$5 2$5 !1$05 12$5 !2$05 !4$%5 0$5

0$%41 !0$1250 0$45%3 !0$150 2$1250 !0$341 !0$%0%3 0$1250





m/ factors

4$5156 he steps 2$

0$116

0$6534 euation $

0$0156

sum 5$3014

e number of dumm' effects 1$3253 uare root of the )ariance 1$1512


r t!)alue

euation #

an effect is

3I6 .&%5

3I6 7 t


30/44



# 5 % & '



" " " ! ! ! " !

" " ! ! ! " ! "

" ! ! ! " ! " "

! ! ! " ! " " !

! ! " ! " " ! "

! " ! " " ! " "

" ! " " ! " " "

! " " ! " " " !

" " ! " " " ! !

" ! " " " ! ! !

! " " " ! ! ! "

! ! ! ! ! ! ! !

36$40 42$25 4%$%5 3#$#5 45$05 3%$0 34$5 42$0

44$55 3%$0 32$10 41$00 35$#0 42$25 46$20 3%$25

%0$#5 %0$#5 %0$#5 %0$#5 %0$#5 %0$#5 %0$#5 %0$#5

!%$15 3$55 16$5 !1$05 #$15 !3$55 !11$45 4$45

!1$35%3 0$5#1 !.&& !0$150 1$5250 !0$5#1 !1$#0%3 0$41





m/ factors

2$3256 he steps 2$0$3501

3$641 euation $

0$5501

sum 6$%65

e number of dumm' effects 1$16#



r t!)alue


31/44

euation #

an effect is

3I6 !.


9 these proteins appear at the top or rather at the bottom of the gel and a correct integration of the pe

ot used for the e)aluation of the :lac*ett!;urman design$

g two tables(


# 5 % & '



" " " ! ! ! " !

" " ! ! ! " ! "

" ! ! ! " ! " "

! ! ! " ! " " !

! ! " ! " " ! "

! " ! " " ! " "

" ! " " ! " " "

! " " ! " " " !" " ! " " " ! !

" ! " " " ! ! !

! " " " ! ! ! "

! ! ! ! ! ! ! !

51$15 56$20 #3$35 32$05 64$60 60$%5 4#$65 6$%5

$40 2$35 35$20 #6$50 63$#5 6$0 %$#0 60$0

12%$55 12%$55 12%$55 12%$55 12%$55 12%$55 12%$55 12%$55

!26$25 !16$15 5%$15 !64$45 0$65 !6$%5 !2#$25 $15

10$23 14$05 15$56 10$6% 16$15 15$21 12$41 13$5

1#$35 14$4 11$3 16$0% 12$# 13$54 15$% 15$1%

-.! !0$4200 3$%250 -5.# 3$3600 1$625 !3$365 !1$6050





m/ factors

3I6 7 t


32/44

11$2%#6 he steps 2$

2$#3

11$3401 euation $

2$560

sum 2%$002#

e number of dumm' effects $000

uare root of the )ariance 2$645#


r t!)alue

euation #

an effect is

3I6 #.5%&



# 5 % & '



" " " ! ! ! " !" " ! ! ! " ! "

" ! ! ! " ! " "

! ! ! " ! " " !

! ! " ! " " ! "

! " ! " " ! " "

" ! " " ! " " "

! " " ! " " " !

" " ! " " " ! !

" ! " " " ! ! !

! " " " ! ! ! "

! ! ! ! ! ! ! !3$4 36$6 2$2 30$2 22$6 25$6 24$% 23$6

22$% 23$ 33$0 30$1 3$6 34$ 35$5 36$

60$2 60$2 60$2 60$2 60$2 60$2 60$2 60$2

14$6 12$# !5$% 0$1 !15 !#$1 !10$ !13$1

$5 6$1 5$4 5$0 5$ 5$1 5$0 5$#

4$6 5$# 6$6 $5 6$3 6$# $1 6$1

!.! 0$1#2 !1$1600 -!.#'&5 !0$616 !1$%200 !2$1400 !0$220%


3I6 7 t


33/44




m/ factors

0$3%03 he steps 2$

3$3124

4$5#6 euation $

0$04%%

sum %$3210

e number of dumm' effects 2$0%03



r t!)alue

euation #

an effect is

3I6 !.!#


3I6 7 t


34/44

response

response809:

1#$5

6$6

#$0

11$5

1$

6$#

5$3

$023$3

6$%

%$#

$4


35/44

$1$ to 2$3$ are summeri7ed in

onsidered as significant at a gi)en if

response

809:

$%

5$0

5$04$2

12$0

26$2

6$4

$#

20$0

6$6

$

$2

2

.Di1-

Documents

Supporting Material_Plakett-Burman Design (1)