For PD-L1, cells were stained with APC-conjugated anti-PD-L1 antibodies (17-5983-42, Thermo Fisher Scientific) according to the manufacturer’s instructions. For uptake assays of FITC-labeled CpG, cells were incubated with 1 μM CpG-A (ODN2216) or CpG-B (ODN2006) for 0-120 minutes. Cells were washed with PBS and analyzed immediately by flow cytometry. Data was acquired on a BD FACSCalibur (BD Biosciences) and analyzed using FlowJo software (version 10).
Apc conjugated anti pd l1 antibody
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom, United States
APC-conjugated anti-PD-L1 antibodies are fluorescent-labeled antibodies that specifically bind to the PD-L1 protein. PD-L1 is an immune checkpoint molecule that plays a role in regulating the immune response. These antibodies can be used in flow cytometry and other immunoassays to detect and analyze PD-L1 expression.
Automatically generated - may contain errors
4 protocols using apc conjugated anti pd l1 antibody
1
PD-L1 and CpG Uptake Assay
2
PD-L1 and CpG Uptake Assay
3
CD24, CD44, and PD-L1 expression in breast cancer cell lines
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HCC1954 and SKBR3 cell variants (1 × 105/well) were seeded in a 6-well plate and allowed to attach overnight. They were then trypsinised, blocked with 10% FCS in PBS and stained with APC-conjugated anti-CD24 (1:75 dilution, eBiosciences, UK) and FITC-conjugated anti-CD44 (1:800 for HCC1954, 1:400 for SKBR3, eBiosciences, UK) or APC-conjugated anti-PD-L1 antibody (1:800 for HCC1954, 1:400 for SKBR3, eBiosciences, UK) for 30 min at 4°C. For EV treatment, cells were seeded at 0.5 × 105 (HCC1954) or 1 × 105 (SKBR3 and EFM192A) cells/well in a 24-well plate and allowed to attach overnight. The following day, cells were washed with serum-free media, fed with media supplemented with extracellular vesicle (EV)-depleted-FBS and treated with EVs for a further 48 hr before staining as above. Staining was assessed in a FACSCanto II flow cytometer, followed by analysis with BD FACSDiva software.
4
Cell Surface Protein Expression Analysis
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The cell surface exposures of CRT, HSP70, HSP90, and human PD-L1 proteins were detected using flowcytometry. Briefly, the cells were treated with various concentrations of CM-EE (0–200 µg/ml) for 72 h and stained with primary antibody for CRT (Cell Signaling, USA), HSP70, or HSP90 (Abcam, United Kingdom) and secondary antibody (Alexa flour 750-conjugated anti-mouse Fc antibody, Abcam, United Kingdom). For PD-L1 expression, cells were treated with CM-EE for 48 h and stained with APC-conjugated anti-PD-L1 antibody (ebioscience, USA). Fluorescence was analyzed using the FACS CantoII flow cytometer and FlowJo software (BD, USA).
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