Tricine-SDS-PAGE是用于分离分子量在1-10kD的肽类用的。
一、 试剂配配制：
1． Low Bis acrylamide（49.5% T, 3% C）
Acylamide 48.0g
Bis 1.5g
Water make up to 100ml
2. High Bis acrylamide (49.5% T, 6% C)
Acrylamide 46.5g
Bis 3.0g
Water make up to100ml
3. Gel Buffer (3M Tris/cl, PH8.45, 0.3% SDS)
SDS 0.3g
Tris 36.4g
Water make up to 100ml
PH to 8.45 with HCL
注: 3M Tris 配制时不容易溶解,因此配制时我配制了2M Tris,配方如下:
Tris 36.4g
SDS 0.3g
Water make up to 150ml
4. Anode (Lower) buffer 阳极缓冲液 (0.2M Tris/cl, PH8.9)
Tris 12.11g
Water make up to 500ml
PH to 8.9 with HCL
5. Cathode (Upper) buffer 阴极缓冲液 (0.1M Tris/cl, 0.1M Tricine, 0.1% SDS, PH 8.25)
Tris 6.06g
Tricine 8.96g
SDS 0.5g
Water make up to 500ml
注:不用调PH值
6. 4×Tricine SDS Sample buffer 4×上样缓冲液
(Final concentration: 0.05M Tris/cl, 4% SDS, 12% glycerol, 200mM DTT, 0.01% Commasie G 250)
8×Tris.cl/SDS PH 6.8 2ml
Glycerol 4.8ml
SDS 1.6g
Commasie Blue G 250 4mg
Water make up to 10ml
注:8×Tris.cl/SDS PH 6.8配方
Tris 6.05g
SDS 0.4g
Water make up to 100ml
PH to 6.8 with HCL
二、 胶的配制
1. 16.5% T 6% C separating gel 30ml
49.5% T 6% C 10ml
Gel buffer 15ml
Glycerol 3.2ml
Water 1.8ml
2. 10% T 3% C spacer gel 30ml
49.5% T 3% C 6.1ml
Gel buffer 15ml
