Natural proteins can be used in measuring intracellular Ca(2+) concentration. As one of the Ca(2+)- regulated photoproteins, aequorin has several advantages in comparison to widely used Ca(2+) fluorescence indicators (e.g., fura-2, indo-1 and fluo-3), including high dynamic range and resistance to motion artefacts. However, incorporation of aequorin into cells remains a challenge. Hypoosmotic shock treatment was optimized and used as a method for loading aequorin into the cytoplasm of follicular lymphoma cells. Measurement of aequorin luminescence in the cells was performed using a luminometer with a sensitive photomultiplier tube and the luminescence intensity was recalculated into intracellular [Ca(2+)]. The value of (0.85 ± 0.52)·10-6 M was found. We show that the optimized method of incorporation was effective for loading aequorin into follicular lymphoma cells in vitro. The cell viability remains high immediately after the procedure. This method can also be used for measuring intracellular Ca(2+) concentration in other types of non-adherent cells.
- MeSH
- aequorin metabolismus MeSH
- aniliny metabolismus MeSH
- folikulární lymfom metabolismus MeSH
- indoly metabolismus MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- vápník metabolismus MeSH
- viabilita buněk fyziologie MeSH
- xantheny metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- izoenzymy fyziologie MeSH
- L-laktátdehydrogenasa fyziologie MeSH
- lidé MeSH
- tvorba protilátek MeSH
- Check Tag
- lidé MeSH
- MeSH
- dieta aterogenní MeSH
- kožní nemoci dietoterapie patologie MeSH
- králíci MeSH
- morčata MeSH
- Check Tag
- králíci MeSH
- morčata MeSH
