1、Western Blotting 所用溶液1.5 M Tris-HCl (pH 8.8) 500 mlTris-base 90.85 gddH2O 400 ml用浓盐酸调pH 至8.8定容至500 ml,高压灭菌,室温保存。1 M Tris-HCl (pH 6.8) 500 mlTris-base 60.57 gddH2O 400 ml用浓盐酸调pH 至6.8定容至500 ml,高压灭菌,室温保存。10%(w/v) AP (过硫酸铵) 20 mlAP 2gddH2O 20 ml分装到 1.5ml离心管,1ml/tube,-20C保存10%(w/v) SDS (十二烷基硫酸钠) 100 mlSD
2、S 10gddH2O 90 ml用磁力搅拌器搅拌直至完全溶解,用水定容至100ml,室温保存。10x Running Buffer (电泳缓冲液) 500 ml1X Running Buffer : 0.025M Tris, 0.25M Glycine, 0.1% SDS10X Running Buffer : 0.25M Tris, 2.5M Glycine, 1% SDS配方:Tris 15.1 g甘氨酸 93.75 gSDS 5.0 gddH2O 400ml,搅拌直至完全溶解后,定容至500ml,室温保存。电泳时10x Running Buffer取100ml,加ddH 2O 900ml
3、,定容至1L,配成1X Running Buffer.1X Running Buffer可反复使用 4-5次。10x Transfer Buffer (转膜缓冲液) 500 ml1X Transfer Buffer: 0.025M Tris, 0.192M Glycine, 0.037% SDS,20% Methanol10X Transfer Buffer: 0.25M Tris, 1.92M Glycine, 0.37% SDS配方:Tris 15.1 g甘氨酸 72 gSDS 1.85 gddH2O 400ml,搅拌直至完全溶解后,定容至500ml,4C保存。转膜时10x Transfer Buffer取100ml,加ddH 2O 500ml,Methanol 200ml,再定容至1L ,配成1X Transfer Buffer,冷却至4C。湿转法1X Running Buffer可反复使用2-3次。10x TBS (缓冲液) 1L (20 mM Tris-HCl, pH 7.5) Tris 24.23 g NaCl 80.06 g ddH2O 800ml pH to 7.5 with HCl. Top up to 1 L. 高压灭菌,室温保存。TBST For 1 L: 100 ml of 10x TBS + 900 ml ddH2O + 1ml Tween20