PC3 and DU145 cells were stained with Brilliant Violet 421™ (BV421) mouse anti-human PD-L1 1:50 (CD274, Clone 29E.2A3, BioLegend, San Diego, CA, USA), PE mouse anti-human HLA-A,B,C 1:100 (Clone W6/32, BioLegend, San Diego, CA, USA), and eFluor™ 780 fixable viability dye 1:10000 (65,086,514, Invitrogen, Waltham, MA, USA). Staining was performed in PBS for 20 min at room temperature. Cells were analyzed using a BD FACSCelesta (Becton Dickinson, Franklin Lakes, New Jersey, USA) and FlowJo software (Tree Star, Ashland, Oregon, USA). The relative mean fluorescence intensity (MFI) was calculated as the MFI of each experimental sample/MFI of vehicle (DMSO) control.
Cd274 clone 29e 2a3
Manufactured by BioLegend
Sourced in United States
CD274, also known as PD-L1, is a cell surface protein that plays a key role in regulating immune responses. The clone 29E.2A3 is a monoclonal antibody that specifically binds to CD274. This antibody can be used in various research applications, such as flow cytometry and immunohistochemistry, to detect and study the expression of CD274 on cells.
Automatically generated - may contain errors
Lab products found in correlation
Efluor 780, by Thermo Fisher Scientific (2 mentions)
Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (2 mentions)
Attune nxt flow cytometer, by Thermo Fisher Scientific (2 mentions)
Lmpo24, by Novus Biologicals (2 mentions)
Abc total compensation beads, by Thermo Fisher Scientific (2 mentions)
Facscelesta, by BD (1 mentions)
Fixable viability dye efluor 780, by Thermo Fisher Scientific (1 mentions)
3 protocols using cd274 clone 29e 2a3
1
Quantification of PD-L1 and HLA-A,B,C Levels
2
Multiparametric Flow Cytometry Analysis
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All stained/fixed samples were acquired on Attune NxT Flow Cytometer (Thermo Fisher Scientific) with necessary internal controls to help assign gates. Fluorescence from multiple antibodies were compensated using AbC Total Compensation beads (Thermo Fisher Scientific, catalog no. A10497). In all experiments, cells were collected and stained with fixable viability dye eFluor780 (1:2000 dilution Life Technologies, catalog no. 65‐0865‐14;) followed by surface staining for PDL1 (CD274; clone 29E.2A3; BioLegend catalog no. 329714; 1:60 dilution) as per the manufacturer's instructions. Cells were then fixed with 4% methanol‐free formaldehyde (Thermo Fisher Scientific, catalog no. 28908) followed by intracellular staining for BZLF1 (Santa Cruz Biotechnology, catalog no. sc‐53904; 1:60 dilution) and LMP1 (clone LMPO24; Novus Biologicals, catalog no. NBP2‐50383; 1:60 dilution) using FoxP3/Transcription factor staining buffer set (eBioscience, catalog no. 5523) as per manufacturer's instructions. Data were analyzed using FlowJo and cumulated using GraphPad PRISM software.
3
Multiparametric Flow Cytometry of PDL1, BZLF1, and LMP1
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