Blotting with BSA
- Blot in 5% mile/ 1X PBST (or 1X TBST) --60'/Rt
- Wash 2X for 5' in 1XPBST
- 10Ab in 1% BSA/1X PBST --60'/Rt
- Wash PBST 15', then 3x for 5' each in 1X PBST
- 20Ab in 1% BSA/1X PBST --60'/Rt (aHRP)--
- Wash 15' with 1X PBST then 3x with PBST for 5' @
- Chemiluminescence: Mix 1:1 NEN reagents. Add to Mb --1'/shaking. Blot excess
- Put between sheets (page protector) and expose film
Dr. DE Koshland, Carnegie Institution of Washington
http://www.ciwemb.edu/labs/koshland/Protocols/Protein/moreblot.html |
