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Methods:Determining intracellular pH

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Contents

Purpose

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Materials Needed

pH-dependent ratiometric GFP clone - pHluorin

Protocol

  1. Clone and express pH-sensitive GFP (pHluorin)
  2. Fluoresence assay (excitation at 410/470m & emission at 520nm)
  3. Determine fluorescence intensity ratio

Links

User notes

References

This paper describes pHluorin. [1]

This paper uses the pHluorin clone to characterize an E. coli Nramp transporter ortholog. [2]

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  1. Miesenböck, G et al. (1998) Visualizing secretion and synaptic transmission with pH-sensitive green fluorescent proteins. Nature 394 192-5 PubMed EcoliWiki page
  2. Courville, P et al. (2004) Determination of transmembrane topology of the Escherichia coli natural resistance-associated macrophage protein (Nramp) ortholog. J. Biol. Chem. 279 3318-26 PubMed EcoliWiki page