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A Simple Flow Cytometric Method to Measure Glucose Uptake and Glucose Transporter Expression for Monocyte Subpopulations in Whole Blood
Published on: August 12, 2016
A flow cytometric screening test for detergent-resistant surface antigens in monocytes
Zsuzsanna Wolf1, Evelyn Orsó, Tobias Werner
1Institute for Clinical Chemistry and Laboratory Medicine, University of Regensburg, Regensburg, Germany.
Summary
This study demonstrates how to identify cell surface proteins associated with lipid rafts in monocytes using flow cytometry. This method efficiently screens raft-association of monocyte antigens in whole blood.
Area of Science:
- Cell Biology
- Immunology
Background:
- Lipid rafts are membrane microdomains crucial for cellular processes.
- Understanding antigen localization in these rafts is key to immune cell function.
Purpose of the Study:
- To investigate the raft-association of surface antigens on human peripheral blood monocytes.
- To evaluate flow cytometry as a method for assessing detergent resistance of monocyte antigens.
Main Methods:
- Utilized flow cytometry to analyze surface antigens on resting and lipopolysaccharide (LPS)-stimulated monocytes.
- Assessed antigen detergent resistance (raft-association) using 0.01% Triton X-100.
Main Results:
- Demonstrated constitutive raft-association for CD14, CD32, and CD55.
- Showed LPS-induced raft-association for CD81.
- Identified CD71 as detergent-soluble (raft-excluded).
Conclusions:
- Flow cytometric detergent insolubility is an effective technique for rapidly screening raft-association of monocyte antigens.
- This method is applicable to whole-blood assays for immunological studies.
