MULTIPARAMETER IMMUNOFLUORESCENCE


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Table of Contents

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Reasons Antibodies do not bind to cells:

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BLOCKING IS IMPORTANT

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variation in gamma 1 myeloma protein binding to macrophages

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DEAD CELLS CAN BE A PROBLEM

ANTIBODIES BIND NON-SPECIFICALLY TO DEAD CELLS

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Evaluating Viability with Ethidium Monoazide

ONE COLOR IMMUNOPHENOTYPING

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Analysis of List Mode Data

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SINGLE COLOR HISTOGRAMS

TWO COLOR PATTERN

THREE COLOR PATTERN

FOUR COLOR SINGLE LASER IMMUNOPHENOTYPING

FOUR COLOR DUAL LASER IMMUNOPHENOTYPING

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VISUALIZING EACH POPULATION

UNDERSTANDING BINARY LOGIC IS USEFUL

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STAINING SOP

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Non-specific Binding to Monocytes

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EFFECT OF BATCH VARIATION

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Author: Carleton C. Stewart, Sigrid J. Stewart, Roswell Park Cancer Institute

Email: stewart@sc3101.med.buffalo.edu

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