Protocol for the dot blot 10-5-09

From OpenWetWare

Jump to: navigation, search
1. wet nitrocellulose with 10ml of 1X TBST for 5minutes
2. make a 2% BSA block (.2g BSA in 10ml 1X TBST)
3. block for 30 minutes
4. wash 3X with TBST - 5minutes/wash
5. add 5ml of 1:500 Ab: TBST, wait 30minutes (on rocker?)
6. wash 3X with TBST - 5minutes/wash
7. leave the last wash on for developing

making 10X TBST: 31.4 g TRIS HCl, 80g NaCl, 10ml Tween-20, pH 7.5, make 1000ml

His HRP Ab --> do 1:500 dilution in 1X TBST

Personal tools