In situ end labelling: effect of proteolytic enzyme pretreatment and hydrochloric acid.
AUTOR(ES)
Carr, N. J.
RESUMO
AIM: To determine the effect of dilute hydrochloric acid on the in situ end labelling (ISEL) reaction, with and without a variety of different proteolytic enzymes. METHODS: Sections of tissue fixed in buffered formalin were pretreated with trypsin, protease XIV (at two different concentrations), or protease XXIV (for two different incubation times), with and without subsequent 1 M hydrochloric acid treatment. The results were compared with those obtained using hydrochloric acid alone, with proteinase K, and pepsin pretreatment, and with no pretreatment. RESULTS: When hydrochloric acid was added to the sections in addition to trypsin, protease XIV, and protease XXIV, there was a significant increase in ISEL reactivity in both apoptotic nuclei and morphologically normal nuclei. Hydrochloric acid alone had no significant effect. CONCLUSIONS: Hydrochloric acid has a distinctive effect on the ISEL reaction that is dependent on prior proteolytic digestion.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=379612Documentos Relacionados
- Simple method for pretreatment of tissue sections for the detection of apoptosis by in situ end-labelling and in situ nick translation
- Alkaline phosphatase for immunocytochemical labelling: problems with endogenous enzyme activity.
- In situ end-labelling, light microscopic assessment and ultrastructure of apoptosis in lung carcinoma.
- Traffic light labelling: traduzindo a rotulagem de alimentos
- Pretreatment with Urea-Hydrochloric Acid Enhances the Isolation of Helicobacter pylori from Contaminated Specimens
