Cell Lysis Buffer

Open Nuclear

Strong
Total Vol.
Stock Conc. Final Conc.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15

dd. H2O
Tris (pH8.0)
NaCl
Na3VO4
NaF
Na4P2O7
EDTA
EGTA
BRIJ
NP-40
Triton X-100
DeOxycholic Acid
SDS
TLA (-20C)
PMSF (-20C)

1M
5M
0.1 M
0.5 M
100 mM
0.5 M
0.5 M
10%
10%
10%
10%
10%
100X
200 mM

50 mM
150 mM
1 mM
10 mM
10 mM
1 mM
1 mM
1%
1%
1%
0.5%
0.1%
1X
2 mM

Medium

Regular

Gentle

Protein-Protein

1000
ul
606
50
30
10
20
100
2
2

1500
ul
909
75
45
15
30
150
3
3

1000
ul
666
50
30
10
20
100
2
2

1500
ul
999
75
45
15
30
150
3
3
0

1000
ul
666
50
30
10
20
100
2
2
100

1500
ul
999
75
45
15
30
150
3
3
150

100

150

100
50
10
10
10

150
75
15
15
15

10
10

15
15

10
10

15
15

1800
ul
1199
90
54
18
36
180
3.6
3.6
180
0
0
0
0
18
18

Nonidet P-40 Lysis Buffer:

1
2
3
4
5
6
7
8

dd. H2O
Tris (pH7.4)
NaCl
NaF
NP-40
DeOxycholic Acid
TLA (-20C)
PMSF (-20C)

Stock Conc.

Total Vol.
Final Conc.

1M
5M
0.5 M
10%
10%
100 X
200 mM

20 mM
150 mM
50 mM
1%
0.5%
1X
1 mM

IP Antibody Dilute Buffer :

1
2
3
4

dd. H2O
Tris (pH7.4)
NaCl
Sodium azide

Stock Conc.

Store at 4C
Total Vol.
Final Conc.

1M
5M
2%

100 mM
150 mM
0.05%

Stock Conc.

Store at 4C
Total Vol.
Final Conc.

1M
5M
0.1 M
0.5 M
0.5 M
10%

50 mM
150 mM
1 mM
1 mM
1 mM
0.1%

IP Wash Buffer :

1
2
3
4
5
6
7

dd. H2O
Tris (pH8.0)
NaCl
Na3VO4
EDTA
EGTA
Triton X-100

1000
ul
685
20
30
100
100
50
10
5

1500
ul
1027.5
30
45
150
150
75
15
7.5

1000
ul
845
100
30
25

100
ml
89.6
5
3
1
0.2
0.2
1

250
ml
224
12.5
7.5
2.5
0.5
0.5
2.5

5000
ul
3425
100
150
500
500
250
50
25

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18

ul
Total loading
Vol. ul
1200
1110
999
900
810
750
660
600
540
510
450
396
330
240
180
150
120
90

1X
6X loading
buffer, ul
200
185
166.5
150
135
125
110
100
90
85
75
66
55
40
30
25
20
15

5%
98% 2MP
ul
61.2
56.6
51.0
45.9
41.3
38.3
33.7
30.6
27.6
26.0
23.0
20.2
16.8
12.2
9.2
7.7
6.1
4.6

ddH2O
ul
938.8
868.4
781.5
704.1
633.7
586.7
516.3
469.4
422.4
399.0
352.0
309.8
258.2
187.8
140.8
117.3
93.9
70.4

Cell Lysis Buffer

NP40

1
2
3
4
5
6
7
8
9
10
11

dd. H2O
Tris (pH8.0)
NaCl
Na3VO4
NaF
Na4P2O7
EDTA
EGTA
NP-40
TLA (-20C)
PMSF (-20C)

Regular
Stock Conc.

Total Vol.
Final Conc.

1M
5M
0.1 M
0.5 M
100 mM
0.5 M
0.5 M
10%
100X
200 mM

50 mM
150 mM
1 mM
10 mM
10 mM
1 mM
1 mM
1%
1X
2 mM

1000
ul
666
50
30
10
20
100
2
2
100
10
10

10ml

1500
ul
999
75
45
15
30
150
3
3
150
15
15

2000
ul
1332
100
60
20
40
200
4
4
200
20
20

2500
ul
1665
125
75
25
50
250
5
5
250
25
25

3000
ul
1998
150
90
30
60
300
6
6
300
30
30

3500
ul
2331
175
105
35
70
350
7
7
350
35
35

4000
ul
2664
200
120
40
80
400
8
8
400
40
40

4500
ul
2997
225
135
45
90
450
9
9
450
45
45

5000
ul
3330
250
150
50
100
500
10
10
500
50
50

5500
ul
3663
275
165
55
110
550
11
11
550
55
55

6000
ul
3996
300
180
60
120
600
12
12
600
60
60

8000
ul
5328
400
240
80
160
800
16
16
800
80
80

1500
ul
909
75
45
15
30
150
3
3
150
75
15
15
15

2000
ul
1212
100
60
20
40
200
4
4
200
100
20
20
20

2500
ul
1515
125
75
25
50
250
5
5
250
125
25
25
25

3000
ul
1818
150
90
30
60
300
6
6
300
150
30
30
30

3500
ul
2121
175
105
35
70
350
7
7
350
175
35
35
35

4000
ul
2424
200
120
40
80
400
8
8
400
200
40
40
40

4500
ul
2727
225
135
45
90
450
9
9
450
225
45
45
45

5000
ul
3030
250
150
50
100
500
10
10
500
250
50
50
50

5500
ul
3333
275
165
55
110
550
11
11
550
275
55
55
55

6000
ul
3636
300
180
60
120
600
12
12
600
300
60
60
60

8000
ul
4848
400
240
80
160
800
16
16
800
400
80
80
80

*TLA & PMSF add before use

RIPA

1
2
3
4
5
6
7
8
9
10
11
12
13

dd. H2O
Tris (pH8.0)
NaCl
Na3VO4
NaF
Na4P2O7
EDTA
EGTA
NP-40
DeOxycholic Acid
SDS
TLA (-20C)
PMSF (-20C)

Strong
Stock Conc.

Total Vol.
Final Conc.

1M
5M
0.1 M
0.5 M
100 mM
0.5 M
0.5 M
10%
10%
10%
100 X
200 mM

50 mM
150 mM
1 mM
10 mM
10 mM
1 mM
1 mM
1%
0.5%
0.1%
1X
2 mM

*TLA & PMSF add before use

1000
ul
606
50
30
10
20
100
2
2
100
50
10
10
10

10ml

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31

Total loading
Vol. ul
48
47
46
45
44
43
42
41
40
39
38
37
36
35
34
33
32
31
30
29
28
27
26
25
24
23
22
21
20
19
18

1X
6X loading
buffer, ul
8.0
7.8
7.7
7.5
7.3
7.2
7.0
6.8
6.7
6.5
6.3
6.2
6.0
5.8
5.7
5.5
5.3
5.2
5.0
4.8
4.7
4.5
4.3
4.2
4.0
3.8
3.7
3.5
3.3
3.2
3.0

5%
98% 2MP
ul
2.4
2.4
2.3
2.3
2.2
2.2
2.1
2.1
2.0
2.0
1.9
1.9
1.8
1.8
1.7
1.7
1.6
1.6
1.5
1.5
1.4
1.4
1.3
1.3
1.2
1.2
1.1
1.1
1.0
1.0
0.9

Protein Vol.
ul
37.6
36.8
36.0
35.2
34.4
33.6
32.9
32.1
31.3
30.5
29.7
28.9
28.2
27.4
26.6
25.8
25.0
24.3
23.5
22.7
21.9
21.1
20.3
19.6
18.8
18.0
17.2
16.4
15.6
14.9
14.1

Mitochondria Lysis Buffer

(1% Chaps Buffer)

Stock Conc.
1
2
3
4
5
6
7
8
9

Ref.

dd. H2O
Tris-HCl (pH 7.4/7.5)
1M
MgCl2
1M
KCl
1M
EDTA
0.5 M
EGTA
0.5 M
Chaps
10%
TLA (-20C)
100X
PMSF (-20C)
200 mM
*TLA & PMSF add before use

Total Vol.
Final Conc.
20 mM
5 mM
137 mM
1 mM
1 mM
1%
1X
2 mM

Nature Cell Biology Vol.6, No. 5, May 2004, 443-450

5000
ul
3670
100
25
685
10
10
500
1ul/100ul
1ul/100ul