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Techniques cellulaires
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Fractionnement cellulaireCentrifugation differentielle
Le principe de fractio!ement cellulaire par centrifugation differentielle consiste a separer les organites selon leur ma!e. Elle permet
l'obtention d'un culot qui se depose au fond du tube et d'obtenir un surnagent. Cela permet de recuperer les fractions.
Les etapes experimentales sont :
- Collecte des cellules
- Homogeneisation a l'aide d'un Potter
- Centrifugation ba"e vite"e pour eliminer les noyaux, les cellules non ca"es
- Recuperation du surnagent
Centrifugation sur gradient de densite
On utilise la force centrifuge pour separer les organites d'un homogenat cellulaire les uns des autres selon leur difference de densite.
Les etapes experimentales sont :
- Collecte de cellules
- Homogeneisation a l'aide d'un Potter
- Centrifugation ba"e vite"e pour eliminer les noyaux, les cellules non ca"es
- Recuperation du surnageant et depot sur un gradient de sucrose
- Centrifugation
- Prelevement par fractions a partir du fond du tube
Western Blot
Le Western Blot permet de detecter la presence d'une proteine d'interet dans l'echantillon
SDS PAGE
Le SDS PAGE permet de separer les proteines en fonction de leur poids moleculaire.
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