Absolute Quantification of Plasma Membrane Receptors Via Quantitative Flow Cytometry

Area of Science:

• Cell Biology
• Biotechnology
• Immunology

Background:

• Plasma membrane receptors are key in cellular signaling and disease diagnostics.
• Measuring receptor abundance is crucial for understanding disease progression.
• Quantitative flow cytometry (qFlow) is a powerful technique for receptor analysis.

Purpose of the Study:

• To detail a quantitative flow cytometry (qFlow) protocol for absolute plasma membrane receptor quantification.
• To establish a standardized method for measuring receptor abundance using phycoerythrin (PE) calibration.
• To provide a foundational protocol for studies involving tyrosine kinase and G-protein-coupled receptors.

Main Methods:

• Utilized quantitative flow cytometry (qFlow) with fluorophore-conjugated antibodies.
• Employed phycoerythrin (PE) as the fluorophore for absolute quantification.
• Developed a detailed protocol for standardized and reproducible receptor measurements.

Main Results:

• Achieved absolute quantification of plasma membrane receptors using qFlow and PE calibration.
• Demonstrated a reproducible method for measuring receptor abundance.
• Established a foundational protocol applicable to various receptor types and sample sources.

Conclusions:

• The described qFlow protocol enables accurate absolute quantification of plasma membrane receptors.
• This method is essential for biomarker discovery and disease diagnostics.
• The protocol supports ongoing research in receptor quantification for tyrosine kinase and G-protein-coupled receptors.