rhPD-1-Fc, anti-PD-L1, IgG1-Fc, and Goat IgG (R&D systems) were solubilized in PBS and added at varying concentrations in 100μL/well to 96-well MaxiSorp plates (ThermoFisher) before overnight incubation at 4°C. Plates were washed with PBS before counted NK-92 cells were added at a concentration of 4 × 104 cells/mL in 200 μL per well in NK-92 media and incubated for 4 hours at 37°C. Degranulation was measured via addition of 2 μg/mL of anti-CD107a (Biolegend) for the duration of the assay. PMA and ionomycin were added to positive control wells at 0.5 μg/mL and 1 μg/mL, respectively, for the duration of the assay.
Rhpd 1 fc
Manufactured by R&D Systems
Sourced in United Kingdom
RhPD-1-Fc is a recombinant fusion protein consisting of the extracellular domain of human Programmed Cell Death 1 (PD-1) and the Fc region of human IgG1. It is produced in a mouse myeloma cell line.
Automatically generated - may contain errors
2 protocols using rhpd 1 fc
1
NK Cell Degranulation Assay with PD-1/PD-L1
2
Osteoclast Differentiation and Modulation
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Osteoclasts were generated from SFMCs, as previously described (32 (link)). Cultures were stimulated with 25 ng/ml recombinant human (rh) macrophage colony-stimulating factor (M-CSF) (Peprotech, London, UK) and 50 ng/ml rhRANKL (Abcam, Cambridge, UK). At each media change, stimulations were added, including rhPD-1Fc (1 μg/ml) (R&D Systems, Abingdon, UK), rhPD-L1Fc (1 μg/ml) (R&D Systems, Abingdon, UK), or anti-RANKL (denosumab, 5 μg/ml). All cultures were evaluated after 21 days for tartrate-resistant acid phosphatase (TRAP) activity (B-Bridge International, CA, USA). Matrix metalloproteinase 9 (MMP-9) was measured by ELISA at days 4, 10, and 19 (R&D Systems).
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