Use of fluorescent Ca2+ dyes with green fluorescent protein and its variants: problems and solutions.
We have studied the degree to which fluorescent Ca(2+) indicator dyes, and green fluorescent protein and its variants, can be used together. We find that the most commonly used fluorescent protein, enhanced green fluorescent protein (EGFP), seriously contaminates fura 2 signals. We suggest two alternative combinations for which there is no detectable contamination of the Ca(2+) indicator signal by the fluorescent protein. Blue fluorescent protein can be used with the Ca(2+) indicator Fura Red; EGFP can be used with the Ca(2+) indicator X-Rhod 1. The use of these combinations will permit the accurate measurement of Ca(2+) signals in cells transfected with fluorescent proteins.
Bolsover
S
S
Department of Physiology, University College London, Gower Street, London WC1E 6BT, UK. s.bolsover@ucl.ac.uk
Ibrahim
O
O
O'luanaigh
N
N
Williams
H
H
Cockcroft
S
S
D000818:Animals; D001572:Benzofurans; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling; D005456:Fluorescent Dyes; D016257:Fura-2; D014644:Genetic Variation; D049452:Green Fluorescent Proteins; D007093:Imidazoles; D008164:Luminescent Proteins / Q000235:genetics; D051381:Rats; D013050:Spectrometry, Fluorescence; D014162:Transfection; D014407:Tumor Cells, Cultured
D001572:Benzofurans; D005456:Fluorescent Dyes; D007093:Imidazoles; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; C082877:fura red; D002118:Calcium; D016257:Fura-2