A Spectrofluorophotometrical Method Based on Fura-2-AM Probe to Determine Cytosolic Ca2+ Level in Pseudomonas syringae Complex Bacterial Cells

Autor: Simone Trabalza, Roberto Buonaurio, Alberto Del Pino, Carlo Palmerini, Harrold van den burg, Chiaraluce Moretti
Jazyk: angličtina
Rok vydání: 2021
Předmět:
Zdroj: Bio-Protocol, Vol 11, Iss 6 (2021)
Druh dokumentu: article
ISSN: 2331-8325
DOI: 10.21769/BioProtoc.3949
Popis: Calcium signaling is an emerging mechanism by which bacteria respond to environmental cues. To measure the intracellular free-calcium concentration in bacterial cells, [Ca2+]i, a simple spectrofluorometric method based on the chemical probe Fura 2-acetoxy methyl ester (Fura 2-AM) is here presented using Pseudomonad bacterial cells. This is an alternative and quantitative method that can be completed in a short period of time with low costs, and it does not require the induction of heterologously expressed protein-based probes like Aequorin. Furthermore, it is possible to verify the properties of membrane channels involved in Ca2+ entry from the extracellular matrix. This method is in particular valuable for measuring [Ca2+]i in the range of 0.1-39.8 µM in small cells like those of prokaryotes.
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