Human CD4+ T cell proliferation was determined by the Click-it EdU detection assay (Thermo Fisher) [60 (link)]. Briefly, 2.5 × 105 CD4+ purified T cells were activated for 72 h in 600 µL RPMI containing 50 µM 5-Ethynyl-2´-deoxyuridine (EdU) and αCD28 (0.05 µg/ml), in sterile 48 well plates precoated with 400 µL of αCD3 (0.5 µg/ml in PBS), and in RPMI containing PMA/Ionomycin. EdU incorporation into DNA from proliferating cells was detected by a fluorescent bioconjugate and flow cytometry. The assay was performed per the manufacturer’s instructions.
Click it edu detection assay
Manufactured by Thermo Fisher Scientific
The Click-iT EdU detection assay is a tool for analyzing cell proliferation. It uses a modified nucleoside, EdU (5-ethynyl-2'-deoxyuridine), which is incorporated into DNA during cell division. The incorporated EdU can then be detected through a copper-catalyzed click reaction, allowing visualization and quantification of proliferating cells.
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CD4+ T Cell Proliferation Assay
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CD4+ T Cell Proliferation Assay
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CD4+ T cell proliferation was determined by the Click‐it EdU detection assay (Thermo Fisher).[42 (link)
] Briefly, 2.5 × 105 CD4+ purified T cells were activated for 72 h in 600 µL cDMEM containing 50 µM 5‐Ethynyl‐2´‐deoxyuridine (EdU) and αCD28 (0.05 µg mL−1), in sterile 48 well plates precoated with 400 µL of αCD3 (0.5 µg mL−1 in PBS). EdU incorporation into DNA from proliferating cells was detected by a fluorescent bioconjugate and flow cytometry. The assay was performed per the manufacturer's instructions.
] Briefly, 2.5 × 105 CD4+ purified T cells were activated for 72 h in 600 µL cDMEM containing 50 µM 5‐Ethynyl‐2´‐deoxyuridine (EdU) and αCD28 (0.05 µg mL−1), in sterile 48 well plates precoated with 400 µL of αCD3 (0.5 µg mL−1 in PBS). EdU incorporation into DNA from proliferating cells was detected by a fluorescent bioconjugate and flow cytometry. The assay was performed per the manufacturer's instructions.
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