Aminopeptidase 345 



3. Transfer to cold (4°C) freshly prepared solution B, agitate: 3-5 

 minutes till blue dye produced. 



4. Wash, 3 changes of cold 0.85% NaCl or water (fixed tissues): 10 

 mintites each. 



5. Motnit in glycerol jelly. 



Burstone Method: (simultaneotis dye and substrate procedure) 



1. Deparaffinize in petroleum ether, 3 changes: 4 minutes each. 



2. Acetone, 2 changes: 4 minutes each, 



3. Acetone-water (1:1): 2 minutes. 



4. Rinse in distilled water: 30 seconds. 



5. Incubate, room temperature: 45-60 minutes. 



6. Rinse in distilled water. 



7. Mount in glycerol jelly. 



results: 

 sites — blue 



comments: 



If decalcification is necessary, Schajoiuicz and Cabrini (1959) say 

 that 2% formic acid and 20% sodium citrate (1:1), pH 5 is satisfac- 

 tory up to 15 days. There is loss of enzyme if versene is tised. 



References: Burstone (1959B); Burton (1954); Goetsch and Reynolds 

 (1951); Gomori (1950A, 1956); Rutenburg and Seligman (1955); and 

 Tandler (1953). 



Aminopeptidase 



Burstone and Folk (1956) 

 FIXATION and embedding: 



1. Cold acetone fixation and paraffin embedding, page 339. 



2. Freeze drying and embedding in \acutun gives better retention of 

 enzyme. 



solutions: 



Substrate and Dye: 



Stock solution of substrate: 



L-leucyl-6-naphthylamide 1.0 gm. 



distilled water 100.0 ml. 



