Apc cy7 anti mouse cd8a

Manufactured by BD

Sourced in United States

APC-Cy7 anti-mouse CD8a is a fluorescently-labeled antibody that specifically binds to the mouse CD8a surface antigen. It can be used in flow cytometry applications to identify and analyze CD8-positive cells.

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2 protocols using apc cy7 anti mouse cd8a

Multiparametric Flow Cytometry Analysis of Immune Cell Subsets

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Flow cytometry was used to characterize the peripheral blood mononuclear cells (PBMCs) and splenocytes. Peripheral blood was collected via retro-orbital bleeding. Splenocytes were collected from spleens that were homogenized by grinding between glass slides. The cells were stained with the following antibodies conjugated to fluorophores: Alexa Fluor 700- anti-mouse CD4 (BD Biosciences, San Jose, CA, United States), PE-Cy5- anti-mouse CD25 (eBioscience, San Diego, CA, United States), PE-CF594- anti-mouse Foxp3 (BD Biosciences), PE- anti-mouse CTLA-4 (eBioscience), FITC-anti-mouse Helios (Biolegend, San Diego, CA, United States), PE-anti-mouse CD11b (BD Biosciences), APC-Cy7 anti-mouse CD8a (BD Biosciences), and Alexa Fluor 700-anti-mouse B220 (eBioscience). The cells were fixed and permeabilized before staining with the eBioscience Foxp3/transcription factor staining buffer set. Flow cytometry was performed on a LSRII flow cytometer (BD Biosciences) and data were analyzed using FlowJo software (FlowJo LLC, Ashland, OR, United States). Gating strategy is included in the Supplementary Figure 1.

Chen A.C., Cai X., Li C., Khoryati L., Gavin M.A, & Miao C.H. (2020). A Treg-Selective IL-2 Mutein Prevents the Formation of Factor VIII Inhibitors in Hemophilia Mice Treated With Factor VIII Gene Therapy. Frontiers in Immunology, 11, 638.

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Immunohistochemistry, Western Blot, and Flow Cytometry Analyses of PD-L1 in Osteosarcoma

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Antibodies for immunohistochemistry were anti‐PD‐L1 (10084‐MB55; Sino Biological), anti‐Ki‐67 (ab16667; Abcam). Antibodies for western blot analyses were anti‐PD‐L1 (ab213524; Abcam), anti‐phospho‐STAT3 (9145; Cell Signaling Technology), anti‐STAT3 (9139; Cell Signaling Technology), and anti‐GAPDH (5174; Cell Signaling Technology). Antibodies for flow cytometry were APC‐anti‐human PD‐L1 (563741; BD Biosciences), BV421‐anti‐mouse PD‐L1 (564716; BD Biosciences), APC‐anti‐human CD3 (555342; BD Biosciences), PE‐anti‐human CD8 (561949; BD Biosciences), APC‐anti‐mouse CD3 (553059; BD Biosciences), PE‐anti‐mouse CD4 (557308; BD Biosciences), APC‐CY7‐anti‐mouse CD8a (561967; BD Biosciences), PE‐CY7‐anti‐mouse CD8a (561097; BD Biosciences). Lymphoprep density gradient centrifugation was purchased from Axis‐Shield (1114546; Axis‐Shield). Neutralizing Abs were anti‐human PD‐L1 neutralizing Ab (16‐5983‐82; eBioscience) and anti‐mouse PD‐L1 neutralizing Ab (16‐5982‐85; eBioscience). STATTIC was purchased from Selleck (S7024). Human recombinant TSP1 (3074‐TH‐050; R&D Systems), mouse recombinant TSP1 (7859‐TH‐050; R&D Systems), human IFN‐γ (C005; Novoprotein), human IL‐6 (C009; Novoprotein), and IL‐10 (CX04; Novoprotein) were used to stimulate osteosarcoma cells.

Liu Z., Wen J., Hu F., Wang J., Hu C, & Zhang W. (2021). Thrombospondin‐1 induced programmed death‐ligand 1‐mediated immunosuppression by activating the STAT3 pathway in osteosarcoma. Cancer Science, 113(2), 432-445.

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