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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
 

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 50C.
2. Prepare stripping buffer.
20 mls: 15 mls ddH2O
1 ml 1 M NaPi, pH 6.5
4 mls 10% SDS
150 l of -mercaptoethanol
Stripping buffer is 50 mM NaPi, pH 6.5, 2% SDS, 0.1M -mercaptoethanol.
3. Place blot in PBSTw at 25C. If blot is dry, wet gradually in PBSTw. The blot must be completely
wet prior to submerging in stripping buffer. Keep blot at 25C 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 50C in Boekel incubator.

  

Source: Aris, John P. - Department of Anatomy and Cell Biology, University of Florida

 

Collections: Biology and Medicine