| Chapter title |
ELISpot and DC-ELISpot Assay to Measure Frequency of Antigen-Specific IFNγ-Secreting Cells.
|
|---|---|
| Chapter number | 8 |
| Book title |
ELISA
|
| Published in |
Methods in molecular biology, January 2015
|
| DOI | 10.1007/978-1-4939-2742-5_8 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-2741-8, 978-1-4939-2742-5
|
| Authors |
Navarrete, Marcelo A, Marcelo A. Navarrete, Navarrete, Marcelo A. |
| Abstract |
ELISpot is a highly sensitive method in immunology to enumerate cells producing a given cytokine. Cells are stimulated in a microtiter plate pre-coated with a specific anti-analyte antibody. In response to the stimulation, cells release cytokines that are bound to the anti-analyte antibody. After a washing step, which removes the cells from the wells, the location of the cytokine-releasing cell is visualized by an enzyme-labeled detection antibody and its corresponding chromogenic substrate. The end result is a set of colored spots, each of which represents an area where a cell secreting the cytokine had been located.Here we describe the standard ELISpot protocol and a variation denominated dendritic cell (DC)-ELISpot for the detection of IFNγ-secreting cells upon stimulation with oligopeptides and protein antigens, respectively. |
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