A Workflow to Achieve Saturation of Fluorophore‐Conjugated Monoclonal Antibodies for Robust Comparison of Biomarker Expression

• Published in: Cytometry. Part A Vol. 107; no. 6; pp. 378 - 389
• Main Authors: Smith, Natalie, McGuire, Helen, Fazekas de St Groth, Barbara
• Format: Journal Article
• Language: English
• Published: Hoboken, USA John Wiley & Sons, Inc 01.06.2025; Wiley Subscription Services, Inc
• Subjects: Animals; Antibodies, Monoclonal - chemistry; Antibodies, Monoclonal - immunology; antibody saturation; Biomarkers; Biomarkers - analysis; CD3 antigen; CD3 Complex - immunology; flow cytometry; Flow Cytometry - methods; Fluorescence; Fluorescent Dyes - chemistry; Humans; Immunoglobulin G; Immunoglobulin G - immunology; Mice; Monoclonal antibodies; Reagents; Robustness; Saturation; Staining; Staining and Labeling - methods; Titration; Workflow; titration; antibody saturation; flow cytometry

ABSTRACT Antibody titration is an important step in every cytometric workflow, with the goal being to determine antibody concentrations that ensure highly reproducible results. When aiming to compare antigen expression between samples using mean or median fluorescence intensity (MFI), reagents shoul...