For flow cytometry (fluorescence-associated cell sorting; FACS), analysis, cells were washed in staining buffer (SB) (2% FBS, 0,1% sodium azide in phosphate-buffered saline) and after a blocking of 10 min with SB+Ab serum 20%, were stained for 30 min with mouse monoclonal antibodies. The antibodies used were: anti-CD16, anti-CD107a, anti EGFR, anti-major histocompatibility complex type I (MHC-I), anti-PD-L1 (Miltenyi Biotec). Stained cells were washed two times, resuspended in SB and then acquired on a FACS ACCURI C6 (BD Biosciences). Analysis was conducted using ACCURI C6 software (BD Biosciences).
Anti pd l1
Manufactured by Miltenyi Biotec
Sourced in United States
Anti-PD-L1 is a laboratory product designed for use in research applications. It functions as an antibody that binds to the PD-L1 protein, which plays a role in immune system regulation. The core function of this product is to facilitate the study of PD-L1 and its interactions, without interpretation or extrapolation on its intended use.
Automatically generated - may contain errors
Lab products found in correlation
Facs accuri c6, by BD (2 mentions)
Accuri c6, by BD (2 mentions)
Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions)
Limulus amebocyte lysate test, by Cambrex (1 mentions)
Anti hla dr, by Immunostep (1 mentions)
Fully human igg4 s228p anti pd 1 receptor blocking monoclonal antibody, by Bristol-Myers Squibb (1 mentions)
Rpmi medium, by Thermo Fisher Scientific (1 mentions)
Anti tim 3, by Miltenyi Biotec (1 mentions)
Anti cd86, by BioLegend (1 mentions)
Anti cd80, by BioLegend (1 mentions)
Anti cd1c, by BioLegend (1 mentions)
Pe labeled anti cd11b, by BioLegend (1 mentions)
Anti cd11c, by BioLegend (1 mentions)
Anti cd83, by BioLegend (1 mentions)
Anti dc sign, by BioLegend (1 mentions)
Facscalibur system, by BD (1 mentions)
Cellquest software, by BD (1 mentions)
4 protocols using anti pd l1
1
Multiparametric Flow Cytometry for Immune Cell Analysis
2
Immunomodulation Assay with PD-1/PD-L1 Inhibitor
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Roswell Park Memorial Institute (RPMI) medium (Invitrogen) was used for the cell cultures. The following antibodies were used: anti-CD14, anti-HLA-DR, anti-CD3 (Immunostep), and anti-PD-L1 (Miltenyi Biotec). The LPS from Salmonella abortus was a kind gift from Dr. Galanos (Max Planck Institute of Immunobiology and Epigenetics). Carboxyfluorescein succinimidyl ester (CFSE) for the proliferation assays was purchased from Thermo Fisher. The lymphocyte stimulus pokeweed (PWD) was purchased from Sigma-Aldrich. To inhibit PD-1/PD-L1 interaction, a fully human IgG4 (S228P) anti-PD-1 receptor-blocking monoclonal antibody was used (Bristol-Myers Squibb). All the reagents used for cell cultures were endotoxin-free, as assayed with the Limulus amebocyte lysate test (Cambrex).
3
FACS Analysis of PBMC Activation Markers
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For flow cytometry (fluorescence-associated cell sorting, FACS) analysis, PBMCs were washed in staining buffer (SB) (2% fetal bovine serum, FBS, 0,1% sodium azide in phosphate-buffered saline) and, after blocking for 10 min with SB plus Ab serum 20%, were stained for 30 min with the following monoclonal antibodies: anti-CD107a, anti-TIM-3 and anti-PD-L1 (Miltenyi Biotec). Stained cells were washed two times, resuspended in SB, acquired on FACS ACCURI C6 and analyzed using ACCURI C6 software (BD Biosciences).
4
Flow Cytometry Analysis of Dendritic Cell Phenotype
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For flow cytometry analysis of DC phenotype we used the following monoclonal antibodies (mAbs): FITC-labeled anti-CD1a (clone: HI149), anti-CD1c (clone: L161), anti-CD14 (clone: 63D3), anti-CD16 (clone: 3G8), and anti-HLA I (clone: W6/32) (all from Biolegend, CA, USA); Alexa Fluor 488-labeled anti-CD4 (clone: RPA-T4) (Biolegend); PE-labeled anti-CD11b (clone: ICRF44), anti-CD11c (clone: 3.9), anti-DC-SIGN (clone: 9E9A8), anti-CD80 (clone: 2D10), anti-CD83 (clone: HB15e), anti-CD86 (clone: BU63) (all from Biolegend), and anti-HLA II (clone: AC122), anti-ILT-3 (clone: REA141), anti-ILT4 (clone: REA184), anti-PDL-1 (clone: REA1197), and anti-FasL (clone: NOK-1) (all from Miltenyi Biotec). Briefly, variously treated DCs were harvested and collected by centrifugation. Antibody was added and the cells were incubated for 15 min in the dark, then washed twice and resuspended in 2% paraformaldehyde (PFA). Samples were analyzed on a FACSCalibur system (Becton Dickinson, Inc.). Data was analyzed with the CellQuest software (BD biosciences).
Endocytosis was monitored by flow cytometry after incubation of DCs with FITC-dextran (1 mg/ml), either on ice or at room temperature, for 1 h. The cells were then washed twice with DPBS and re-suspended in 2% PFA for future analysis.
Endocytosis was monitored by flow cytometry after incubation of DCs with FITC-dextran (1 mg/ml), either on ice or at room temperature, for 1 h. The cells were then washed twice with DPBS and re-suspended in 2% PFA for future analysis.
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