Biology Reference
In-Depth Information
Microdomains
2500
L of 10% Triton X-100
m
100
L of 0.5 M EDTA, pH 8.0
m
500
L of fresh 0.1 M Na
3
VO
4
m
500
L of 100 mM PMSF
m
L of 10 mg/mL aprotinin
42.6 mL of ddH
2
O
MBS buffer
(25 mM MES pH 6.5, 150 mM NaCl)
50
m
FOR 300 ML
7.5 mL of 1 M MES buffer, pH 6.5
15 mL of 3 M NaCl
277.5 mL of ddH
2
O
40 mL MBS buffer)
35% (w/v) sucrose
: Dilute 85% sucrose in MBS with 1 mM EDTA and 1 mM
Na
3
VO
4
4 SAMPLES (21 ML)
8.65 mL of 85% sucrose
210
85% (w/v) sucrose
(64 g sucrose
þ
L of 0.1 M Na
3
VO
4
42
m
L of 0.5 M EDTA
12.098 mL of MBS
m
6 SAMPLES (31 ML)
12.77 mL of 85% sucrose
310
L of 0.1 M Na
3
VO
4
m
62
L of 0.5 M EDTA
17.858 mL of MBS
m
5% (w/v) sucrose
: Dilute 85% sucrose in MBS with 1 mM EDTA and 1 mM
Na
3
VO
4
4 SAMPLES (21 ML)
1.24 mL of 85% sucrose
210
L of 0.1 M Na
3
VO
4
m
42
L of 0.5 M EDTA
19.508 mL of MBS
m
6 SAMPLES (31 ML)
1.83 mL of 85% sucrose
310
m
L of 0.1 M Na
3
VO
4
L of 0.5 M EDTA
28.798 mL of MBS
Note
: Mix sucrose mixtures at 4
C, rotating for 2 min.
62
m
2
Sample buffer
Dilute 4
sample buffer (refer to
Section 2.1.2
) with DIG lysis buffer (1:1)
For example, 100
Lof4
sample buffer
þ
100
L of DIG lysis buffer
m
m
