The BP cell line was treated with 20 ng/ml IFNγ in DMEM with 10% FBS for 24 hours. Cells were trypsinized to generate a single-cell suspension and stained with either anti-PD-L1 (10F.9G2; BioLegend) or MHC-I (AF6-88.5; BD Biosciences). Samples were analyzed on an LSR II (BD Biosciences) and data were analyzed with FlowJo software (TreeStar).
Mhc 1 af6 88.5
Manufactured by BD
MHC-I (AF6-88.5) is a lab equipment product that is used for the detection and analysis of major histocompatibility complex class I (MHC-I) molecules. It functions as a tool for researchers to study the expression and characteristics of MHC-I proteins in various cell types and biological systems.
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Lab products found in correlation
Anti pd l1 10f 9g2, by BioLegend (1 mentions)
Oxldl, by Thermo Fisher Scientific (1 mentions)
Zombie fixable viability dye, by BioLegend (1 mentions)
Lsrfortessa, by BD (1 mentions)
Nile red, by Thermo Fisher Scientific (1 mentions)
Cholesterol assay kit, by Abcam (1 mentions)
Mhcii, by BD (1 mentions)
Facsdiva software, by Tree Star (1 mentions)
2 protocols using mhc 1 af6 88.5
1
IFNγ-Induced PD-L1 and MHC-I Expression
2
Flow Cytometry Analysis of Lipid Metabolism
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Flow cytometry staining were performed in FACS buffer (PBS 1% BSA, 1% FCS) using the following anti-mouse antibodies from Biolegend unless specified otherwise (at a dilution of 1:200): CD1d (1B1), CD11b (M1/70), CD11c (N418), CCR7 (4B12), CD45.1 (A20), CD45.2 (104), CD24 (M1/69), CD40 (3/23), CD117 (2B8), CD86 (GL-1), TLR4 (SA15-21), MHC-I (AF6-88.5, BD Biosciences), MHC-II (M5/114.15.2), CD36 (HM36), MSR1 (1F8C33), CD80 (16-10A1), CD69 (H1.2 F). Dead cells were excluded from the analyses by staining with DAPI or Zombie fixable viability dye (Biolegend). Flow cytometry data were collected on an LSR-II or LSRFortessa (both from BD Biosciences) using FACSDiva Software and were analysed with FlowJo software (TreeStar).
Cholesterol staining with Filipin III was performed using Cholesterol Assay Kit (Abcam) and lipid staining was performed using Nile Red (ThermoFisher) according to the manufacturer’s instructions. For CD36 internalization, pMACs were treated with 2 μg/ml αCD1d (19G11, InVivoMab) or isotype control for 1 h, prior to stimulation with 250 μg/ml oxLDL (ThermoFisher) for a further 1 h.
Cholesterol staining with Filipin III was performed using Cholesterol Assay Kit (Abcam) and lipid staining was performed using Nile Red (ThermoFisher) according to the manufacturer’s instructions. For CD36 internalization, pMACs were treated with 2 μg/ml αCD1d (19G11, InVivoMab) or isotype control for 1 h, prior to stimulation with 250 μg/ml oxLDL (ThermoFisher) for a further 1 h.
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