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Elispot for Rookies (and Experts Too)

Elispot for Rookies (and Experts Too)


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International Edition


About the Book

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.


About the Author: 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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Product Details
  • ISBN-13: 9783319832609
  • Publisher: Springer
  • Publisher Imprint: Springer
  • Height: 234 mm
  • No of Pages: 93
  • Series Title: Techniques in Life Science and Biomedicine for the Non-Exper
  • Weight: 209 gr
  • ISBN-10: 3319832603
  • Publisher Date: 05 Jul 2018
  • Binding: Paperback
  • Language: English
  • Returnable: N
  • Spine Width: 6 mm
  • Width: 156 mm


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