Anti cd8 pe

Manufactured by Beckman Coulter

Sourced in United States

The Anti-CD8-PE is a fluorescently labeled antibody that specifically binds to the CD8 surface antigen. It is designed for use in flow cytometry applications to identify and quantify CD8-positive cells within a sample.

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CD8-PE (7 citations) Anti-CD8 (6 citations) PE/Cy7 anti-human CD8 (2 citations)

6 protocols using anti cd8 pe

Profiling Immune Cell Subsets in Cancer

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Frequency of T cell subsets, macrophage subsets, and myeloid-derived suppressor cells (MDSC) in the peripheral blood was analyzed via flow cytometry. PBMCs procured from pre-therapy and cycle 4 day 1 blood draws were stained with anti-CD3-V450, anti-CD8-PE, anti-CD4-FITC, anti-Foxp3-PE, anti-CD56-APC, anti-CD16-APC Cy7, anti-NKG2C, anti-CD33-APC, anti-HLA-DR-PECy7, anti-CD11b-PE, anti-CD14-V450, anti-CD15-FITC, anti-CD80-FITC, anti-CD1630-PE, anti-CD206-APC and/or anti-CD69-PE-Texas Red (Beckman Coulter, Brea, CA). Immune cell subsets were defined as follows: CD4 T cells CD3⁺/CD4⁺, CD8 T cells CD3⁺/CD8⁺, Treg CD3⁺/CD4⁺/Foxp3⁺, M1 macrophages CD14⁺/CD80⁺/CD163⁻/CD206⁺, M2 macrophages CD14⁺/CD80/CD163⁺/CD206⁺, NK cells CD3⁻/CD56⁺/CD16⁺, granulocytic MDSC CD33⁺/HLA-DR⁻/CD11b⁺/CD15⁺, monocytic MDSC CD33⁺/HLA-DR⁻/CD11b⁺/CD14⁺. Data was acquired using a LSRII flow cytometer (BD Biosciences, San Jose, CA).

McMichael E.L., Benner B., Atwal L.S., Courtney N.B., Mo X., Davis M.E., Campbell A.R., Duggan M.C., Williams K., Martin K., Levine K.M., Olaverria Salavaggione G.N., Noel T., Ganju A., Uppati S., Paul B., Olencki T., Teknos T.N., Savvides P., Tridandapani S., Byrd J.C., Caligiuri M.A., Liu S.V, & Carson WE I.I.I. (2019). A phase I/II trial of cetuximab in combination with interleukin-12 administered to patients with unresectable primary or recurrent head and neck squamous cell carcinoma. Clinical cancer research : an official journal of the American Association for Cancer Research, 25(16), 4955-4965.

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Comprehensive Immune Cell Profiling

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The following mAbs were used for flow cytometry: anti-CD3 PerCP, anti-CD4 FITC, anti-CD4 PerCP, anti-CD4 PE-Cy7, anti-CD8 PErCP, anti-CD8 APC-Cy7, anti-CD28 FITC, anti-CD95 APC, anti-CD195 (anti-CCR5) PE, anti-Ki67 PE, anti-HLA-DR APC, and anti-HLA-DR FITC (BD Biosciences). Additionally, annexin V PE was purchased from BD Biosciences, anti-CD8 APC and anti-CD8 PE were obtained from Beckman Coulter (CA, USA), and anti-CD38 FITC was purchased from StemCell Technologies (Vancouver, Canada). Flow cytometry data were acquired on a FACSAria II (BD Biosciences). All data were analyzed using FlowJo software (Tree Star, Ashland, OR, USA).

Zhan X.Y., Wang N., Liu G., Qin L., Xu W., Zhao S., Qin L, & Chen X. (2014). Plasmodium infection reduces the volume of the viral reservoir in SIV-infected rhesus macaques receiving antiretroviral therapy. Retrovirology, 11, 112.

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Immunomodulatory Effects of Curcumin in Mice

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Curcumin, hematoxylin, Freund's incomplete adjuvant, mouse TNF-α and IFN-γ were purchased from Sigma-Aldrich (Deisenhofen, Germany). The monoclonal FAPα antibody was from Abcam, and the monoclonal IDO antibody was from Chemicon. The monoclonal anti-vimentin antibody, polyclonal anti-β-actin and secondary HRP-conjugated anti-mouse antibody were from Cell Signaling Technology (MA, USA). The anti-CD8-PE and anti-IFN-γ-FITC were from Beckman Coulter, and IL-2 and Brefeldin A were from BioLegend.

Jiang G.M., Xie W.Y., Wang H.S., Du J., Wu B.P., Xu W., Liu H.F., Xiao P., Liu Z.G., Li H.Y., Liu S.Q., Yin W.J., Zhang Q.G., Liang J.P, & Huang H.J. (2015). Curcumin combined with FAPαc vaccine elicits effective antitumor response by targeting indolamine-2,3-dioxygenase and inhibiting EMT induced by TNF-α in melanoma. Oncotarget, 6(28), 25932-25942.

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Quantification of Serum Biomarkers and Immune Cell Phenotypes

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Blood was centrifuged and serum was stored in -80 °C until assayed. Levels of biomarkers were quantified by using commercially available kits of enzyme immunosorbent assays from Bio-Techne (R&D Systems, Minneapolis, MN) according to the manufacturer’s instructions. The lower limits of detection were: 16 pg/ml for tumor necrosis factor-alpha (TNFα); 40 pg/ml for IL-6; 31 pg/ml for IL-10; 62 pg/ml for IL-38; 156 pg/ml for IFNγ; and 313 p/ml for platelet-derived growth factor (PDGF)-A.
White blood cells were incubated for 15 minutes in the dark with the monoclonal antibodies anti-CD14 FITC, anti-HLA-DR-PE, anti-CD45 PC5 (Beckman Coulter, Marseille, France). White blood cells were also incubated for 15 minutes in the dark with anti-CD3 FITC, anti-CD4 FITC and anti-CD19 FITC (fluorescein isothiocyanate, emission 525nm, Beckman Coulter); with anti-CD4 PE, anti-CD8 PE, and anti-CD(16+56) PE (phycoerythrin, emission 575nm, Beckman Coulter); and with anti-CD45 PC5 (emission 667nm, Beckman Coulter). Fluorospheres (Beckman Coulter) were used for the determination of absolute counts. Cells were analyzed after running through the CYTOMICS FC500 flow cytometer (Beckman Coulter Co, Miami, Florida). Isotypic IgG controls stained also with anti-CD45 were used for each patient. Results of HLA-DR on CD14/CD45-cells were expressed as mean fluorescence intensity (MFI).

Bacharaki D., Karagiannis M., Giannakopoulos P., Papachristou E., Divanis D., Sardeli A., Petrou D., Nikolopoulos P., Bratsiakou A., Zoi V., Piliouras N., Damoraki G., Liakopoulos V., Goumenos D, & Giamarellos-Bourboulis E.J. (2023). Immune responses of patients on maintenance hemodialysis after infection by SARS-CoV-2: a prospective observational cohort study. BMC Infectious Diseases, 23, 581.

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Assessing PBMC Proliferation upon Activation

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PBMCs (4x10⁵) from FAP and healthy subjects were stimulated in a 96-well plate coated sequentially with anti-mouse IgG (4 μg/ml; Southern Biotech), then with anti-CD3 antibodies (5 μg/ml; UCHT1, Biolegend). Soluble anti-CD28 antibody (2 μg/ml; Beckman Coulter) was added to the medium and cells were incubated for 72 h at 37°C. In indicated experiments, PBMCs were labeled with 5 μM of CellTrace™ violet cell proliferation kit (Invitrogen) previous to stimulation, for assessment of cell proliferation. Three days post stimulation, cells were stained with Zombie NIR™ Fixable viability kit (1/1000; Biolegend), next with anti-CD3-APC (1/40; clone UCHT-1; eBiosciences), anti-CD4-PE-Vio770 (1/100; clone rea623; Miltenyi Biotec), anti-CD8-PE (1/10; clone Leu-2a; Beckman Coulter). FACS analyses were performed as above.

Cuche C., Mastrogiovanni M., Juzans M., Laude H., Ungeheuer M.N., Krentzel D., Gariboldi M.I., Scott-Algara D., Madec M., Goyard S., Floch C., Chauveau-Le Friec G., Lafaye P., Renaudat C., Le Bidan M., Micallef C., Schmutz S., Mella S., Novault S., Hasan M., Duffy D., Di Bartolo V, & Alcover A. (2023). T cell migration and effector function differences in familial adenomatous polyposis patients with APC gene mutations. Frontiers in Immunology, 14, 1163466.

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Peripheral Blood Immunophenotyping by FACS

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2ml of heparinized peripheral blood was drawn from each patient. Then 100 μl blood was incubated in the dark with primary antibody at 4°C for 15 min. After hemolysis for 10 min, samples were centrifuged for 10 min at 1500 rpm at room temperature, and then washed twice in PBS and subjected to flowcytometric analysis (Becton-Dickinson, Franklin Lakes, NJ). Primary antibodies included: anti-CD4-FITC, anti-CD8-PE, anti-CD3-PerCP (Becton-Dickinson), anti-CD4-FITC (Beckman-Coulter), anti-CD25-PE (Beckman-Coulter), anti-CD28-FITC (Beckman-Coulter), anti-CD8-PE (Beckman-Coulter), anti-CD3-FITC .

Zhao Y.J., Jiang N., Song Q.K., Wu J.P., Song Y.G., Zhang H.M., Chen F., Zhou L., Wang X.L., Zhou X.N., Yang H.B., Ren J, & Lyerly H.K. (2015). Continuous DC-CIK infusions restore CD8+ cellular immunity, physical activity and improve clinical efficacy in advanced cancer patients unresponsive to conventional treatments. Asian Pacific journal of cancer prevention : APJCP, 16(6).

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