RESUMO
The demonstration of prophenoloxidase after electrophoresis is based on its activation by sodium dodecyl sulfate (SDS) or sodium oleate and staining the activated phenoloxidase with dopamine and 3-methyl-2-benzothiazolinone hydrazone hydrochloride (MBTH). A rapid method is presented for demonstrating the presence of activated phenoloxidase using polyacrylamide gel electrophoresis followed by staining in the presence of SDS or sodium oleate.
Assuntos
Aracnídeos/metabolismo , Catecol Oxidase/análise , Precursores Enzimáticos/análise , Animais , Benzotiazóis , Catecol Oxidase/sangue , Catecol Oxidase/isolamento & purificação , Dopamina , Eletroforese em Gel de Poliacrilamida , Ativação Enzimática/efeitos dos fármacos , Precursores Enzimáticos/sangue , Precursores Enzimáticos/isolamento & purificação , Hidrazonas , Ácidos Oleicos/farmacologia , Dodecilsulfato de Sódio/farmacologia , Coloração e Rotulagem , TiazóisRESUMO
Prophenoloxidase has been successfully extracted in a stable condition from the Hemocytes of Periplaneta americana using cane sugar saline solution. Other media used to extract prophenoloxidase were found not suitable. Among the activators used (trypsin, chymotrypsin, sodium oleate and SDS), trypsin activated the proenzyme maximum. Activation during electrophoresis resulted in the dissociation of proenzyme in to two sub-units. The enzyme has also been localised by the agarose gel electrophoresis. The proenzyme was stable up to 50 degrees C and precipitated at 60 degrees C and above. The activated enzyme showed maximum activity at pH 7.0. Preliminary attempt has also been made to precipitate the enzyme protein against the antibody raised in rabbit.
Assuntos
Catecol Oxidase/metabolismo , Precursores Enzimáticos/metabolismo , Hemolinfa/enzimologia , Periplaneta/enzimologia , Animais , Cromatografia em Gel , Eletroforese em Gel de Ágar , Eletroforese em Gel de Poliacrilamida , Ativação Enzimática/fisiologia , Estabilidade Enzimática/fisiologia , Temperatura Alta , Concentração de Íons de Hidrogênio , Imunodifusão , Cloreto de SódioRESUMO
Lyo- and desmo-glycogen were estimated in relation to growth and maturity of proglottids of a cestode, Moniezia benedeni. Total and lyo-glycogen show an increasing trend from immature to mature region (i.e., from region 1 to 6) and the increased level seen in the 6th region is maintained up to the 9th region with a fall in the 10th (gravid) region. These results indicate that mature proglottids store more carbohydrate than immature and gravid proglottids. Desmo-glycogen showed no variation in different regions of the strobila. The percentage of lyo-glycogen to total carbohydrate reveals that most of the carbohydrate (85-93%) is stored in a labile form.