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Author:   Sylvia Janetzki
Publisher:   Springer International Publishing AG
Imprint:   Springer International Publishing AG
Edition:   1st ed. 2016
Dimensions:   Width: 15.50cm , Height: 0.80cm , Length: 23.50cm
Weight:   0.454kg
ISBN:  

9783319452937

ISBN 10:   3319452932
Pages:   93
Publication Date:   21 November 2016
Audience:   Professional and scholarly ,  Professional & Vocational
Format:   Hardback
Publisher's Status:   Active
Availability:   Manufactured on demand  
We will order this item for you from a manufactured on demand supplier.

Table of Contents

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.

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Author Information

Sylvia Janetzki is the CEO of the ZellNet Consulting, whose services cover all aspects of Elispot, including troubleshooting, review of protocol, set-up for clinical trial monitoring, certification services, evaluation of plates, and training procedures.

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