Titlebook: Elispot for Rookies (and Experts Too); Sylvia Janetzki Book 2016 Springer International Publishing Switzerland 2016 B-cell Assay.Elispot.F

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Important Assay Details,Conducting an Elispot assay is a multistep process. The main steps are:

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Reagents and Materials,The choice of the Elispot plate can dramatically influence the final results of an Elispot experiment. Membrane-based 96-well Elispot plates are the optimal choice for Elispot. The membrane fibers look like a tightly woven web of polymers that offers a large surface for protein binding (Fig. 3.1).

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Planning the Experiment,A good Elispot experiment starts with a well-thought-through plan of execution. The quality of an experiment and its results are determined by the controls it includes. Here is a summary of important controls:

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Quality Control,Throughout the book we have touched upon various important controls, for the sample (apoptosis and viability, see Sect. 4.6), and for the assay (negative controls, sample-specific internal controls, nonspecific positive controls, external controls, see Sect. 5.1).

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Troubleshooting,This book has been structured in a way that it provides detailed information on choices about how to run an optimized Elispot assay. Throughout the book examples are given for what works and what does not and for good and suboptimal outcomes, and recommendations on how to prevent suboptimal outcomes and the reasoning behind it are provided.

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