Chapter title |
Determination of Peroxisomal pH in Living Mammalian Cells Using pHRed
|
---|---|
Chapter number | 16 |
Book title |
Peroxisomes
|
Published in |
Methods in molecular biology, April 2017
|
DOI | 10.1007/978-1-4939-6937-1_16 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6935-7, 978-1-4939-6937-1
|
Authors |
Godinho, Luis F., Schrader, Michael, Luis F. Godinho, Michael Schrader |
Editors |
Michael Schrader |
Abstract |
Organelle pH homeostasis is crucial for maintaining proper cellular function. The nature of the peroxisomal pH remains somewhat controversial, with several studies reporting conflicting results. Here, we describe in detail a rapid and accurate method for the measurement of peroxisomal pH, using the pHRed sensor protein and confocal microscopy of living mammalian cells. pHRed, a ratiometric sensor of pH, is targeted to the peroxisomes by virtue of a C-terminal targeting sequence. The probe has a maximum fluorescence emission at 610 nm while exhibiting dual excitation peaks at 440 and 585 nm, allowing for ratiometric imaging and determination of intracellular pH in live cell microscopy. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 13 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Researcher | 4 | 31% |
Student > Ph. D. Student | 2 | 15% |
Unspecified | 1 | 8% |
Professor | 1 | 8% |
Student > Doctoral Student | 1 | 8% |
Other | 2 | 15% |
Unknown | 2 | 15% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 5 | 38% |
Chemistry | 3 | 23% |
Agricultural and Biological Sciences | 2 | 15% |
Unspecified | 1 | 8% |
Unknown | 2 | 15% |