Apc conjugated anti pd 1

Manufactured by BioLegend

APC-conjugated anti-PD-1 is a lab equipment product from BioLegend. It is a fluorescently-labeled antibody that binds to the PD-1 (Programmed Cell Death 1) receptor on cells.

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Anti-PD-1 (51 citations) Anti-PD-1-APC (18 citations) APC-PD-1 (4 citations) APC-conjugated anti-human PD-1 (3 citations) APC-conjugated anti-human PD-1 antibody (2 citations)

4 protocols using apc conjugated anti pd 1

Measuring CAR Expression on T Cells

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To measure CAR expression, cells were first suspended in 100 μL of cold buffer (PBS containing 0.5% BSA and 2 mM EDTA) supplemented with 2 μg of goat IgG (Jackson Immunoresearch, West Grove, PA, USA) and incubated on ice. CD19-FLAG CAR-T cells were stained with 2 μL of PE- or Alexa Fluor 488-conjugated anti-FLAG (Biolegend), whereas BCMA CAR-T cells were first stained with 0.4 μg of BCMA-huFc protein (Acro Biosystems, Newark, DE, USA) and then stained with 1 μL of PE- or Alexa Fluor 488-conjugated goat anti-human IgG (Jackson Immunoresearch). Cells were co-stained with either FITC-conjugated anti-CD4 and APC-conjugated anti-CD8; PE-conjugated anti-CD27 and APC-conjugated CD45RO; or APC-conjugated anti-PD-1 (all from Biolegend). Dead cells were identified with 7-aminoactinomycin D (7-AAD, BioLegend). The cells were rinsed with 3 mL of buffer, then suspended in buffer and acquired on a FACSCalibur (BD Biosciences, San Jose, CA, USA). Gating strategies are shown in Supplemental Figure S1.

Berahovich R., Liu X., Zhou H., Tsadik E., Xu S., Golubovskaya V, & Wu L. (2019). Hypoxia Selectively Impairs CAR-T Cells In Vitro. Cancers, 11(5), 602.

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Comprehensive Immune Cell Profiling

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For analysis of surface markers, single-cell suspensions were stained with following fluorochrome-conjugated antibodies following established guidelines (61 (link)): FITC-conjugated anti-CD45.1 (Biolegend, 110705), PerCP-Cyanine5.5-conjugated anti-CD8 (Biolegend, 110733), Pacific Blue-conjugated anti-CD8 (Biolegend, 100725), PE-conjugated anti-CD69 (Biolegend, 104508), FITC-conjugated TIM3 (Genetex, GTX54055), APC-conjugated anti-LAG3 (Biolegend, 125210), APC-conjugated anti-PD-1 (Biolegend, 135210), APC-conjugated anti-IL-2 (Biolegend, 503809), APC-conjugated anti-CD25 (Biolegend, 101909), FITC-conjugated anti-IFN-γ (Biolegend, 505805), APC-conjugated anti-TNF-α (Biolegend, 506307), PE-conjugated anti-H-2 K(b)/SIINFEKL Tetramer (Helixgen, HG08T14028), BV421-conjugated KLRG1 (Biolegend, 138413), BV605-CD127 (Biolegend, 135025), and APC-Cy7-conjugated anti-CD62L (Biolegend, 104427), For intracellular staining, cells were first fixed with IC Fixation Buffer (eBioscience 00-8222-49) and permeabilized with Permeabilization Buffer (Invitrogen, 00-8333-56). Flow cytometry was performed using Accuri C6 or Sony ID7000 system and analyzed with FlowJo software.

Zhang H., Yang Z., Yuan W., Liu J., Luo X., Zhang Q., Li Y., Chen J., Zhou Y., Lv J., Zhou N., Ma J., Tang K, & Huang B. (2024). Sustained AhR activity programs memory fate of early effector CD8+ T cells. Proceedings of the National Academy of Sciences of the United States of America, 121(11), e2317658121.

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Flow Cytometric Analysis of Tfh Subsets and Plasma Cells

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We isolated peripheral blood mononuclear cells (PBMCs) from the blood samples using density-gradient centrifugation on Ficoll-Paque. Single-cell suspensions were treated with fixable viability stain 510 (BD Bioscience, 564406) for half an hour, followed by staining with the following antibodies to distinguish the Tfh subsets: FITC- conjugated anti-CD45RA (BioLegend, N418), PE-conjugated anti-CCR6 (BioLegend, G034E3), PerCP/Cy5.5-conjugated anti-CXCR5 (BioLegend, J252D4), PE/Cy7-conjugated anti-CXCR3 (BioLegend, G025H7), APC-conjugated anti-PD-1 (BioLegend, EH12.2H7), and APC/Cy7- conjugated anti-CD4 (BioLegend, RPA-T4). For the detection of circulating plasma cells, PBMCs were stained with FITC-conjugated CD27 (BioLegend, M-T271), PE-conjugated IgD (BioLegend, W18340F), PerCP/Cy5.5-conjugated anti-CD19 (BioLegend, HIB19), and APC-conjugated CD38 (BioLegend, HB-7), followed by the treatment with fixable viability stain 510. Cell acquisition was performed using a CantoII cytometer (BD Bioscience). Data were analyzed with Diva (BD) software.

Mao M., Xu S., Lin L., Dong D., Xue M., He S, & Cai G. (2022). Impact of Corticosteroids on the Proportions of Circulating Tfh Cell Subsets in Patients With Systemic Lupus Erythematous. Frontiers in Medicine, 9, 949334.

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Multicolor Flow Cytometry Analysis

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Cell suspensions from spleens were prepared by grinding tissue through sterile wire mesh and stained with a panel of uorescent-conjugated antibodies including PerCP/Cy5.5 conjugated anti-B220 (Biolegend, 103236), FITC-conjugated anti-FAS (BD Pharmingen, 554257), eFluor 660-conjugated anti-GL7 (Invitrogen, 50-5902-82), PerCP/Cy5.5 conjugated anti-CD4 (Biolegend, 100434 ), PE-conjugated anti-CXCR5 (Biolegend, 145504), and APC-conjugated anti-PD1 (Biolegend, 109112). Samples were run on a FACS Calibur (BD), and the data were analyzed with FlowJo software (TreeStar, Portland, OR).

Xing Y., Guo W., Wu M., Xie J., Huang D., Hu P., Zhou M., Zhang L., Zhang Q., Wang P., Wang X., Wang G., Wu H., Zhou C., Chen Y., Liu M., Yi Z, & Sun Z. (2022). An orally available small molecule BCL6 inhibitor effectively suppresses diffuse large B cell lymphoma cells growth in vitro and in vivo. Cancer letters, 529.

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