Elispot for Rookies (and Experts Too) /
| Autor principal: | |
|---|---|
| Autor Corporativo: | |
| Formato: | eBook |
| Lenguaje: | English |
| Publicado: |
Cham :
Springer International Publishing : Imprint: Springer,
2016.
|
| Edición: | 1st ed. 2016. |
| Colección: | Techniques in Life Science and Biomedicine for the Non-Expert,
|
| Materias: |
Tabla de Contenidos:
- Preface
- Overview
- Historic Overview
- Principles of Elispot
- Modifications
- Simplifications
- Coating Step
- Detection Step
- Adaptations
- Detection of specific antibodies
- Detection of multiple analytes in one well
- Dual color Elispot
- Fluorospot
- Summary
- Important Assay Details
- Reagents and Materials
- Elispot plates
- Antibody choices
- Enzymatic conjugates and substrates
- Fluorescent dyes
- Culture media
- Blocking buffers
- Washing buffers
- Important laboratory supplies
- Pipettes
- Filters
- Other supplies
- Automated Elispot reader
- Sample preparation
- Overview
- Whole blood and PBMC
- Blood draw and anticoagulants
- PBMC isolation
- Time frame between blood draw and PBMC isolation
- Immune cells obtained from tissue
- Freezing of cells
- Thawing of cells
- Cell counting and viability
- Overnight resting
- Isolation of sub-populations of cells
- Expansion of cells
- Planning the experiment
- Controls
- Negative controls
- Positive controls
- External controls
- Plate layout
- Timing
- The Elispot assay
- Coating
- Blocking
- Effector cell preparation
- Stimulant preparation
- Peptides and peptide pools
- Proteins and lysate
- Antigen presenters
- Control stimulants
- Reference samples
- Plating out cells and stimulants
- Incubation
- Removal of cells
- Spot development
- Enumeration of spots
- Data analysis
- Quality control
- Reporting guidelines
- Troubleshooting
- References.