| Chapter title |
Measurement of Export to the Cytosol in Dendritic Cells Using a Cytofluorimetry-Based Assay.
|
|---|---|
| Chapter number | 13 |
| Book title |
Dendritic Cell Protocols
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-3606-9_13 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-3604-5, 978-1-4939-3606-9
|
| Authors |
Omar I. Vivar, Joao G. Magalhaes, Sebastian Amigorena |
| Editors |
Elodie Segura, Nobuyuki Onai |
| Abstract |
The presentation of exogenous antigens on MHC class I molecules, known as cross-presentation, is a key function of dendritic cells (DCs). Cross-presentation via the cytosolic pathway involves antigen export from endocytic compartments to the cytosol. We have recently developed a cytofluorimetry-based assay to examine the kinetics and the efficiency of antigen export to the cytosol in DC populations. In this assay, DCs are loaded with a FRET-sensitive cytosolic substrate of β-lactamase, CCF4. Following uptake of β-lactamase by the DCs, the enzyme undergoes export to the cytosol leading to cleavage of the FRET dye. This cleavage and switch of fluorescence are analyzed by flow cytometry, allowing a quantitative measurement of this event. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 10 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 4 | 40% |
| Other | 1 | 10% |
| Student > Master | 1 | 10% |
| Researcher | 1 | 10% |
| Professor > Associate Professor | 1 | 10% |
| Other | 0 | 0% |
| Unknown | 2 | 20% |
| Readers by discipline | Count | As % |
|---|---|---|
| Immunology and Microbiology | 3 | 30% |
| Agricultural and Biological Sciences | 2 | 20% |
| Biochemistry, Genetics and Molecular Biology | 1 | 10% |
| Chemistry | 1 | 10% |
| Unknown | 3 | 30% |