| | |
Summary: 51
India Ink Staining
1. Place wet or dried blot in a small volume of PBSTw. Place blot on surface of liquid to allow slow and
even wetting.
2. Add India Ink dropwise while swirling buffer (up to 0.1% final concentration).
3. Gently agitate 1 hour to overnight.
4. Destain with PBSTw.
Stripping Blot for Reprobing with Antibody
1. Preheat Boekel incubator to 50°C.
2. Prepare stripping buffer.
20 mls: 15 mls ddH2O
1 ml 1 M NaPi, pH 6.5
4 mls 10% SDS
150 µl of ß-mercaptoethanol
· Stripping buffer is 50 mM NaPi, pH 6.5, 2% SDS, 0.1M ß-mercaptoethanol.
3. Place blot in PBSTw at 25°C. If blot is dry, wet gradually in PBSTw. The blot must be completely
wet prior to submerging in stripping buffer. Keep blot at 25°C on bench until next step.
4. Submerge wet blot in stripping solution in a tightly sealed plastic tray. If the tray does not seal
tightly you will know.
5. Incubate blot in stripping buffer for 60 minutes at 50°C in Boekel incubator.
|
