Pd l1 pe

Manufactured by BD

Sourced in United States

PD-L1-PE is a laboratory reagent used for the detection and quantification of the programmed death-ligand 1 (PD-L1) protein in biological samples. It is a fluorescently-labeled antibody that binds specifically to the PD-L1 protein, allowing for its identification and measurement in various research and analytical applications.

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Lab products found in correlation

Facscanto 2, by BD (7 mentions) Cd13 pe cy7, by BD (3 mentions) Hla abc apc, by BD (3 mentions) Cd31 fitc, by BD (3 mentions) Cd90 apc, by R&D Systems (3 mentions) Cd73 pe, by BD (3 mentions) Cd3 percp cy5, by BD (2 mentions) Cd4 fitc, by BD (2 mentions) Cd45 apc cy7, by BD (2 mentions) Cd8 apc, by BD (2 mentions) Hla dr percp, by BD (2 mentions) Cd11c apc, by BD (2 mentions) Cd105 fitc, by R&D Systems (2 mentions) Facsdiva software, by BD (2 mentions) Gallios flow cytometer, by Beckman Coulter (2 mentions) Cd4 fitc, by BioLegend (2 mentions) Cd45 bv510, by BD (2 mentions) Cd8 pe, by BD (2 mentions) Facscalibur, by BD (2 mentions) Cd8 fitc, by BD (2 mentions)

20 protocols using pd l1 pe

Flow Cytometry Analysis of Tumor-Derived Cells

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All antibodies used for flow cytometry were purchased from BD Pharmingen and included CD3–PerCP-Cy5.5 (Cat. number 561108), CD4-FITC (Cat. number 553047), CD8-APC (Cat. number 553035), CD8-BV510 (Cat. number 563068), CD45–APC-Cy7 (Cat. number 557659), CD45-PE (Cat. number 553081), CD90.2-FITC (Cat. number 553003), CD326 (EpCAM)-APC (Cat. number 563478), FoxP3-PE (Cat. number 560408), and PD-L1–PE (Cat. number 558091). One million cells were stained with 1 μg of each fluorochrome-conjugated antibody and analyzed using a FACS Canto II (Becton Dickinson). Cell surface staining was performed by incubating tumor-derived cell populations with selected antibodies on ice in the dark for 30 minutes in PBS and 0.5% FBS (HyClone GE Healthcare Life Sciences). Intracellular staining of FoxP3 was performed using a mouse FoxP3 fixation and permeabilization kit (BD Pharmingen, cat. number 560409). Non-viable cells were excluded from further flow analysis using a Live/Dead Fixable Violet Dead Cell Stain Kit (ThermoFisher, cat. number L34955). Data were analyzed using FlowJo version 10.1r7 (FlowJo, LLC).

Zhao F., Evans K., Xiao C., DeVito N., Theivanthiran B., Holtzhausen A., Siska P.J., Blobe G.C, & Hanks B.A. (2018). Stromal Fibroblasts Mediate Anti–PD-1 Resistance via MMP-9 and Dictate TGFβ Inhibitor Sequencing in Melanoma. Cancer immunology research, 6(12), 1459-1471.

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Phenotypic Profiling of Stimulated MSCs

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Unstimulated and IFNγ-stimulated MSC were incubated with mouse-anti-human monoclonal antibodies against CD13-PE-Cy7; CD45-APC; HLA-DR-PERCP; HLA-ABC-APC; CD31-FITC; CD73-PE; PD-L1-PE (all BD Biosciences, San Jose, CA, USA); and CD90-APC (R&D Systems, Abingdon, UK) at room temperature in the absence of light for 30 min. After two washes with FACS Flow, flow cytometric analysis was performed using FACSCANTO-II with KALUZA Software (BD, San Jose, CA, USA).

Tejeda-Mora H., Leon L.G., Demmers J., Baan C.C., Reinders M.E., Bleck B., Lombardo E., Merino A, & Hoogduijn M.J. (2021). Proteomic Analysis of Mesenchymal Stromal Cell-Derived Extracellular Vesicles and Reconstructed Membrane Particles. International Journal of Molecular Sciences, 22(23), 12935.

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Comprehensive Immune Cell Profiling

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The following antibodies were used: mTOR mAb (7C10), phospho-S6 (Ser235/236) (D57.2.2E)-APC, phospho-p70 S6 kinase (Thr389) (108D2) mAb, phosphor-AKT (Thr308) (C31E5E) mAb, phospho-Akt (Ser473) (D9E) mAb, GFP (D5.1) mAb, SHP-2 (D50F2) mAb (Cell Signaling Technology); APC-conjugated Donkey-anti-Rabbit IgG (Jackson ImmunoResearch); CD3e (145-2C11)-FITC, CD4 (RM4-5)-PE-Cy7, Vβ4 (KT4)-PE, CD11c-APC, CD11b-PE-Cy7, B220-FITC, CD19-PE-Cy7 and PD-L1-PE (BD Biosciences); F4/80-PE and PD-1-PE-Cy7 (BioLegend); CD71-APC and CD98-PE, CD90.1 (Thy-1.1)-PE, CXCR3-APC, CD80-PE, CD86-PE and PD-L2-PE (eBioscience); PD-1 (J43) Ab (Novus); T-bet-FITC and SH-PTP2 (C-18) (Santa Cruz Biotechnology).

Chen W., Wan X., Ukah T.K., Miller M.M., Barik S., Cattin-Roy A.N, & Zaghouani H. (2016). Antigen-Specific Immune Modulation Targets mTORC1 Function to Drive Chemokine Receptor-Mediated T Cell Tolerance. Journal of immunology (Baltimore, Md. : 1950), 197(9), 3554-3565.

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Immunophenotypic Analysis of AT-MSCs

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Unstimulated and IFNγ-stimulated AT-MSC were trypsinized and washed with FACS Flow (BD Biosciences, San Jose, CA). Cell suspensions were incubated with mouse-antihuman monoclonal antibodies against CD13-PE-Cy7; HLA-DR-PERCP; HLA-ABC-APC; CD31-FITC; CD73-PE; PD-L1-PE (all BD Biosciences); CD90-APC and CD105-FITC (R&D Systems, Abingdon, UK) at room temperature in the absence of light for 30 min. After two washes with FACS Flow, flow cytometric analysis was performed using FACSCANTO-II with FACSDIVA Software (BD Biosciences).

Gonçalves F.D., Luk F., Korevaar S.S., Bouzid R., Paz A.H., López-Iglesias C., Baan C.C., Merino A, & Hoogduijn M.J. (2017). Membrane particles generated from mesenchymal stromal cells modulate immune responses by selective targeting of pro-inflammatory monocytes. Scientific Reports, 7, 12100.

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Monocyte response to microparticle stimuli

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CD14⁺ cells were purified from PBMC using auto-MACS Pro by positive-selection. Monocyte purity was measured by flow cytometry after staining with mouse-antihuman monoclonal antibodies against CD14-PerCP (BD Biosciences) and CD3-PacBlue (BD Biosciences). Isolated CD14⁺ monocytes (2 × 10⁵ cells/200 µl) were cultured in RPMI 1640 medium (Life Technologies), supplemented with 10% FBS and 1% P/S. Monocytes were cultured with MP, or MPγ at different ratios (1:10,000, 1:40,000, 1:80,000) in polypropylene tubes. After 24 h of incubation, monocytes were collected for PCR analysis or flow cytometry after staining with CD14-PacBlue, CD3-PerCP, CD16-FITC, PD-L1-PE and CD90-APC (all BD Biosciences).

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Analyzing PD-L1 Expression on Monocytes

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2 ml of residual peripheral venous blood was obtained on admission from blood samples for clinical use, stored at 4 in a ethylenediamine tetraacetic acid (EDTA) tube and flow cytometry analysis was performed within 2 hours. PD-L1-PE (Cat# 560176) and CD14-APC-H7 (Cat# 560180) and their respective isotype controls recommended were purchased from BD Pharmingen (San Jose, CA, USA). 100 µl of whole blood sample was incubated with anti-CD14 and anti-PD-L1 antibodies for 20 minutes. Red blood cells were lysed. Anti-CD14 and anti-PD-L1 labeled cells and their isotypes labeled cells were obtained and run on a Gallios Flow Cytometer (Beckman Coulter, Inc.). Kaluza analysis version 1.5a (Beckman Coulter, Inc.) software was used for data analysis. Gating protocols of PD-L1 on CD14+ monocytes are shown in Figure 1.

Lu Y., Wang G, & Li C. (2021). Expression of peripheral monocytic programmed death ligand-1 in severe sepsis combined with HBV-related cirrhosis. A pilot observational study. Central-European Journal of Immunology, 46(2), 217-224.

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Comprehensive Immune Cell Profiling

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Extracellular surface marker staining was performed with IgG1-FITC, IgG2a-FITC, IgG1-PE, αCD25-FITC, αCD40-PE, αCD69-PE, αCD70-PE, αCD80-FITC, αCD83-PE, αCD86-FITC, and PD-L1-PE (all from BD Biosciences, Heidelberg, Germany); IgG3-PE (eBioscience, Frankfurt, Germany); and αCCR7-FITC (R&D Systems, Minneapolis, MN, USA) for 30 min at 4 °C in PBS supplemented with 1% FCS and 0.02% sodium azide (Merck, Darmstadt, Germany). The cells were analyzed using a FACScan cytofluorometer equipped with CellQuest software (BD, Heidelberg, Germany). Analysis was performed with the FCS Express software (De Novo Software, Glendale, CA, USA). Specific MFIs were calculated by subtracting the background MFI obtained with the isotype controls.

Hoyer S., Eberlein V., Schuler G., Berking C., Heinzerling L., Schaft N, & Dörrie J. (2021). BRAF and MEK Inhibitors Affect Dendritic-Cell Maturation and T-Cell Stimulation. International Journal of Molecular Sciences, 22(21), 11951.

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Multiparametric Analysis of Tumor-Infiltrating Immune Cells

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Tumors tissues were harvested, and single-cell homogenates were prepared by using a mouse tumor dissociation kit (Miltenyi Biotec) and passed through a 70-mm strainer before the red blood cells were lysed. Surface markers were stained with the following antibodies at 4 °C for 30 min: CD45-BV510 (563891, BD), CD3-PerCP-Cy5.5 (551163, BD), CD4-FITC (100406, BioLegend), CD8-Alexa Fluor 700 (557959, BD), CD69-PE/Cy7 (104512, BioLegend), CD44-BV650 (103049, BioLegend), PD-1-PE-CF594 (562523, BD) and PD-L1-PE (558091, BD). For forkhead box P3 (FOXP3) staining, a transcription factor buffer set (00-5523-00, eBioscience) was used.
For DCs staining, following antibodies were used: CD11c-APC (17-0114-82, eBioscience), MHC I-PE (12-5958-82, eBioscience) and CD86-eFluor 450 (48-0862-82, eBioscience).

Liu Y., Cai J., Liu W., Lin Y., Guo L., Liu X., Qin Z., Xu C., Zhang Y., Su X., Deng K., Yan G, & Liang J. (2020). Intravenous injection of the oncolytic virus M1 awakens antitumor T cells and overcomes resistance to checkpoint blockade. Cell Death & Disease, 11(12), 1062.

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Dendritic Cell Differentiation and Maturation

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CD14+ monocytes were isolated from peripheral blood mononuclear cells of a normal donor post Ficoll (GE Healthcare) separation of lymphocytes using the Miltenyi positive isolation kit (Miltenyi Biotec, Bergisch Gladbach, Germany). Counted cells were resuspended in a 24-well plate at 0.5 × 10⁶/mL in complete media supplemented with IL-4 (100 ng/mL; Peprotech, Rocky Hill, NJ) and granulocyte-macrophage colony-stimulating factor (50 ng/mL, R&D systems). Between days 4 and 6 of cell culture, cells were matured with lipopolysaccharide (100 ng/mL; Sigma–Aldrich, St. Louis, MO). Maturation of the dendritic cells was assessed by staining for major histocompatibility complex class II (FITC; BD Biosciences), B7-2 (CD86 PE; BD Biosciences), and PD-L1 (PE; BD Biosciences).

Malm I.J., Bruno T.C., Fu J., Zeng Q., Taube J.M., Westra W., Pardoll D., Drake C.G, & Kim Y.J. (2014). Expression profile and in vitro blockade of programmed death-1 in human papillomavirus–negative head and neck squamous cell carcinoma. Head & neck, 37(8), 1088-1095.

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Multi-parameter Flow Cytometry Analysis

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Single cells were incubated with mouse Fc receptor blocking reagent (Miltenyi) for 20 min at 4°C, washed, and stained with fluorochrome-conjugated antibodies for 20 min at 4°C. Cells were acquired using a Gallios flow cytometer (Beckman Coulter).
The following antibodies were used (from Miltenyi unless otherwise noted): CD45-Vioblue, clone REA737; CD45-PerCP-Vio770, clone REA737; CD11b-PerCP-Vio700, clone REA592; CD11c-PErCP-Vio700, clone REA754; CD11c-PE, clone REA754; F4/80-PE, clone REA126; XCR1-APC, clone REA707; MHCII-Vioblue, clone REA813; H2K^b/D^b-APC, clone REA932; CD3-PerCP-Vio700, clone REA641; CD4-FITC, clone REA604; CD8-PE, clone REA 601; CD8-APC, clone REA601; PD1-APC, clone J43 (BD); CD86-VioBright, clone REA1190; CD86-PE, clone REA1190; CD172α-APC-Vio-770 (Sirpα), clone REA1201; CD223-FITC (LAG-3), clone C9B7W; NKp46-PE, clone REA815, PD-L1-PE, clone MIH5 (BD).

Moreo E., Uranga S., Picó A., Gómez A.B., Nardelli-Haefliger D., del Fresno C., Murillo I., Puentes E., Rodríguez E., Vales-Gómez M., Pardo J., Sancho D., Martín C, & Aguilo N. (2022). Novel intravesical bacterial immunotherapy induces rejection of BCG-unresponsive established bladder tumors. Journal for Immunotherapy of Cancer, 10(7), e004325.

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