Proteolysis and lectin histochemistry.
Lectin binding to formalin-fixed, paraffin-embedded tissue can often be enhanced by pre-treatment of the sections with proteolytic enzymes. However, the pattern of staining may be profoundly influenced by the type of enzyme preparation which is used. Sites of binding of thirteen different lectins to murine ovary and thyroid gland were studied after exposure of tissue sections to crude trypsin, purified trypsin, purified alpha-chymotrypsin, pepsin, protease VII, papain, bromelain, thermolysin or elastase. With most lectins, the results obtained were similar regardless of which enzyme was used for proteolytic digestion. However, the pattern of binding of soy bean lectin to the ovary and of concanavalin A and common pea lectin to the thyroid gland was highly dependent upon the enzyme used to pre-treat the sections. In both tissues, the staining pattern seen in untreated frozen sections was similar to that found in formalin-fixed, paraffin-embedded material digested with purified trypsin, but was different from that observed after exposure of processed sections to crude trypsin. The location of binding sites after treatment of paraffin sections with chymotrypsin was the same as that after digestion with crude trypsin. Results obtained after the use of other proteolytic enzymes varied according to the tissue being studied. These findings imply that the effect of treatment with crude trypsin is due to contaminating chymotrypsin, and demonstrate that the use of purified trypsin may have advantages over other proteolytic enzymes in lectin histochemistry. The observations may also apply to other related cytochemical techniques such as immunocytochemistry.
D000818:Animals; D005260:Female; D005404:Fixatives; D005557:Formaldehyde; D005615:Freezing; D006651:Histocytochemistry / Q000379:methods*; D037102:Lectins; D051379:Mice; D010053:Ovary / Q000166:cytology / Q000378:metabolism; D010232:Paraffin; D010447:Peptide Hydrolases / Q000378:metabolism*; D013194:Staining and Labeling; D013961:Thyroid Gland / Q000166:cytology / Q000378:metabolism
D005404:Fixatives; D037102:Lectins; D005557:Formaldehyde; D010232:Paraffin; D010447:Peptide Hydrolases