Elimination of the non-specific binding of avidin to tissue sections.
A simple procedure is described for eliminating non-specific staining with avidin-peroxidase conjugates. Murine ovaries were embedded in either paraffin wax or epoxy resin and, after blocking endogenous peroxidase activity, were treated with 10 micrograms/ml biotinylated Pisum sativum agglutinin. Avidin-peroxidase conjugates (5 micrograms/ml), diluted in standard 0.05 M tris-buffered saline, pH 7.6, containing 0.139 M NaCl, produced considerable background coloration and intense mast cell staining in controls without the lectin. This background diminished as the ionic strength of the buffer was raised. At 0.125 M Tris-buffered saline (containing 0.347 M NaCl) the background was completely unstained, with elimination of all binding to mast cells and only minimal loss of specific lectin binding.
D001360:Avidin / Q000378:metabolism*; D002021:Buffers; D006651:Histocytochemistry / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes; D009994:Osmolar Concentration; D009195:Peroxidase; D010544:Peroxidases; D011485:Protein Binding
D002021:Buffers; D007527:Isoenzymes; D001360:Avidin; D010544:Peroxidases; D009195:Peroxidase