Protocol for high-throughput compound screening using flow cytometry in THP-1 cells
| Autor: | Reza Beheshti Zavareh, Stephan H. Spangenberg, Luke L. Lairson |
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| Rok vydání: | 2021 |
| Předmět: |
Science (General)
Cell Survival THP-1 Cells High-throughput screening Immunology Drug Evaluation Preclinical Antineoplastic Agents Cellular level General Biochemistry Genetics and Molecular Biology Flow cytometry Q1-390 medicine Protocol Humans THP1 cell line Throughput (business) General Immunology and Microbiology medicine.diagnostic_test Chemistry General Neuroscience Cell-based assays Membrane Proteins Flow Cytometry Small molecule Cell biology High-Throughput Screening Assays Cell culture Monocytic leukemia Flow cytometry/mass cytometry |
| Zdroj: | STAR Protocols STAR Protocols, Vol 2, Iss 2, Pp 100400-(2021) |
| ISSN: | 2666-1667 |
| Popis: | Summary Flow cytometry is a valuable method for analyzing protein expressions at the single cell level but can be difficult to apply to large numbers of samples. This protocol provides instructions to perform a high-throughput small molecule screen using flow cytometry analysis of THP-1 cells, a human monocytic leukemia cell line. We describe a methodology for identifying compounds that regulate PD-L1 surface expression in IFN-γ-stimulated cells, which has been successfully used to screen a collection of ∼200,000 compounds. For complete details on the use and execution of this protocol, please refer to Zavareh et al. (2020). Graphical Abstract Highlights • Protocol for high-throughput screening of compounds using flow cytometry • Designed for quantification of cell surface expression of proteins in THP-1 cells • This protocol has been used to identify modulators of PD-L1 expression Flow cytometry is a valuable method for analyzing protein expressions at the single cell level but can be difficult to apply to large numbers of samples. This protocol provides instructions to perform a high-throughput small molecule screen using flow cytometry analysis of THP-1 cells, a human monocytic leukemia cell line. We describe a methodology for identifying compounds that regulate PD-L1 surface expression in IFN-γ-stimulated cells, which has been successfully used to screen a collection of ∼200,000 compounds. |
| Databáze: | OpenAIRE |
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