MACSprep™ PBMC Isolation Kit, human

Protocol tips

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Heparinised blood samples were used fresh within 4 h, and peripheral blood mononuclear cells (PBMCs) were separated from blood using Ficoll–Hypaque or NycoPrep.

Upstream tips
Heparinised blood samples were used fresh within 4 h, and peripheral blood mononuclear cells (PBMCs) were separated from blood using Ficoll–Hypaque or NycoPrep.

Publication protocol

Heparinised blood samples were used fresh within 4 h, and peripheral blood mononuclear cells (PBMCs) were separated from blood using Ficoll–Hypaque or NycoPrep. Following this, more refined PBMCs were obtained via MACSprep™ PBMC Isolation Kit. To verify the effect of Wnt5a neutralising antibodies or recombinant human Wnt11 on the suppression of cytokine secretion and NF-κB activation, PBMCs isolated from SARS-CoV-2 patients were incubated with the Wnt5a antibody (20 μg·mL−1) or recombinant human Wnt11 (10 ng·mL−1) for 6 h. The supernatant was used for analysis of cytokines ELISA, and lysate was used for NF-κB activity analysis by an ELISA-based NF-κB family transcription factor assay kit (43296; Active Motif). All experiments were performed independently at least three times. Statistically significant differences were determined using unpaired t-test. Prism software was used for statistical analyses.

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