Western上样缓冲液配制整理
发布时间:2020-04-04 18:45:58
发布时间:2020-04-04 18:45:58
● 6×SDS样品缓冲液
4×Tris·Cl/SDS,PH6.8 (0.28mol/L) | 7ml | |
甘油[30%(V/V)] | 3.0ml(3.8g) | 甘油密度为1.2613g/cm3 (20/4℃) |
SDS [1%(m/V)] | 1g | |
溴酚蓝[0.0012%(m/V)] | 1.2mg | |
DTT (0.5mol/L) | 0.93g | |
如果需要,加去离子蒸馏水至10ml;等量分装成0.5ml并于-70℃贮存
● 8×Tris·Cl/SDS,PH6.8
Tris-base(0.5mol/L) | 6.05g | 40ml水中溶解后以1mol/L盐酸调至PH6.8,补加至100ml。 |
HCl | 调至6.8 | 0.45um滤膜过滤,再加0.4gSDS |
SDS[0.4%(m/V)] | 0.4g | 4℃可保存1月 |
●丽春红S染色液
| 0.2% | |||
SDS Loading buffer | 2× | 10ml | 4× | 10ml |
Tris-HCl PH=6.8 | 0.125 M | 1.25ml(1mol/L) | 0.25 M | 2.5ml(1mol/L) |
SDS | 4% | 0.4g或者 (10%SDS4ml) | 8% | 0.8g |
2-ME2 | 5% | 10% | ||
DTT3 | 0.15 M | 1.5ml(1mol/L) | 0.3 M | 3ml(1mol/L) |
Glycerol | 20% | 2ml | 30% | 3ml |
Bromphenol blue | .01% | 0.001g | .02% | 0.002g |
ddH2O | 1.25ml | 1.5ml | ||
1. Prepared using Tris base, pH adjusted with HCl.
2. If 2-ME is used, omit DTT.
3. If DTT is used, omit 2-ME.