Electrophoretic detection of chitinase isoenzymes using the PhastSystem.
Electrophoresis
; 14(1-2): 165-7, 1993.
Article
em En
| MEDLINE
| ID: mdl-8462510
ABSTRACT
Zymographic assays are described for the detection of chitinase isoenzymes following isoelectric focusing. Method 1 used a polyacrylamide overlay gel containing glycol chitin. Following hydrolysis, visualisation of isoenzymes was achieved by fluorescent counter-staining of the gel with Fluorescent Brightener 28. Method 2 used a cellulose acetate membrane overlay which incorporates 4-methylumbelliferyl substrates, hydrolysis of which released a fluorescent product (4-methylumbelliferone). The pIs of chitinase isoenzymes were estimated by using coloured pI markers. A significant time advantage was obtained over previous methods. The method was used to demonstrate chitinase activity in control rat lungs and in rat lungs infected with the pathogen Pneumocystis carinii.
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Coleções:
01-internacional
Contexto em Saúde:
4_TD
/
6_ODS3_enfermedades_notrasmisibles
Base de dados:
MEDLINE
Assunto principal:
Pneumonia por Pneumocystis
/
Quitinases
/
Focalização Isoelétrica
/
Isoenzimas
/
Pulmão
Tipo de estudo:
Diagnostic_studies
Limite:
Animals
Idioma:
En
Revista:
Electrophoresis
Ano de publicação:
1993
Tipo de documento:
Article