Using of chicken antibodies for metallothionein detection in human blood serum and cadmium-treated tumour cell lines after dot- and electroblotting

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00064203%3A_____%2F09%3A5392" target="_blank" >RIV/00064203:_____/09:5392 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216208:11130/09:5392

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Using of chicken antibodies for metallothionein detection in human blood serum and cadmium-treated tumour cell lines after dot- and electroblotting

Popis výsledku v původním jazyce

Metallothionein (MT) is a protein playing role in homeostasis of metal ions. Its relation with formation and progression of tumors is discussed. Here, we propose a new methodological approach for visualization of MT on PVDF membranes after dot- and electroblotting by using a commercial monoclonal antibody and polyclonalchicken antibodies. The optimized procedure was as follows. We dotted sample on membrane and let it to dry. Then, we blocked the membrane with BSA. After that, the membrane was incubatedin chicken primary antibody, washed, and incubated in secondary antibody conjugated with horseradish peroxidase. To visualize, we used 3-aminoethyl-9-carbazole. Under these conditions, we estimated detection limit as 3 pg of MT per 1 microL. The optimalapproach was further utilized for detection of MT level in 2 human fibroblast lines and in serum obtained from children with medulloblastoma. The results were in good agreement with differential pulse voltammetry ?Brdicka reaction.

Název v anglickém jazyce

Using of chicken antibodies for metallothionein detection in human blood serum and cadmium-treated tumour cell lines after dot- and electroblotting

Popis výsledku anglicky

Metallothionein (MT) is a protein playing role in homeostasis of metal ions. Its relation with formation and progression of tumors is discussed. Here, we propose a new methodological approach for visualization of MT on PVDF membranes after dot- and electroblotting by using a commercial monoclonal antibody and polyclonalchicken antibodies. The optimized procedure was as follows. We dotted sample on membrane and let it to dry. Then, we blocked the membrane with BSA. After that, the membrane was incubatedin chicken primary antibody, washed, and incubated in secondary antibody conjugated with horseradish peroxidase. To visualize, we used 3-aminoethyl-9-carbazole. Under these conditions, we estimated detection limit as 3 pg of MT per 1 microL. The optimalapproach was further utilized for detection of MT level in 2 human fibroblast lines and in serum obtained from children with medulloblastoma. The results were in good agreement with differential pulse voltammetry ?Brdicka reaction.

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

CB - Analytická chemie, separace

OECD FORD obor

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Projekt

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Návaznosti

V - Vyzkumna aktivita podporovana z jinych verejnych zdroju

Rok uplatnění

2009

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Electrophoresis

ISSN

0173-0835

e-ISSN

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Svazek periodika

30

Číslo periodika v rámci svazku

21

Stát vydavatele periodika

DE - Spolková republika Německo

Počet stran výsledku

10

Strana od-do

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Kód UT WoS článku

000272126000011

EID výsledku v databázi Scopus

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