Flow-cytometry analysis was performed on a BD Biosciences LSR-II cytometer equipped with UV, violet, blue, and red lasers. Doublets and dead cells were excluded by forward scatter, side scatter, and use of the AF350 LIVE/DEAD fixable violet dead cell stain (Thermo Fisher, #L34964, 1:1000). B cells were tested for CD19 (BioLegend, #302243 Brilliant Violet 605 anti-human CD19 antibody, 1:200), CD3 (BioLegend, #317329, Brilliant Violet 785 anti-human CD3 antibody, 1:200), and CD38 (BD, #551400, PerCP-Cy5.5 mouse anti-human CD38, 1:200). SupT1 cells were tested for CD8 (BD, #560662, PerCP-Cy5.5 mouse anti-human CD8, 1:100). Cells were fixed with the Cytofix/Cytopermreagent and washed in Perm/Wash buffer (BD). Cells were then permeabilized using Permeabilization Plus Buffer (BD, #561651). B cells were subsequently stained intracellularly for IgG (BioLegend, #410707, PE anti-human IgG Fc antibody, 1:200) and IgM (BioLegend, #314532PE/Cyanine7 anti-human IgM antibody, 1:200), and SupT1 cells were tested for p24 (Beckman Coulter, #6604667, HIV-1 core antigen-RD1, KC57, 1:100).
Percp cy5.5 mouse anti human cd8
Manufactured by BD
Sourced in United Kingdom
PerCP-Cy5.5 mouse anti-human CD8 is a fluorochrome-conjugated monoclonal antibody that binds to the CD8 antigen expressed on the surface of human T cells. It is used in flow cytometry applications for the identification and enumeration of CD8-positive cell populations.
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2 protocols using percp cy5.5 mouse anti human cd8
1
Flow Cytometry Analysis of Immune Cells
2
Quantification of Plasma Antibody Responses
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To assess compliance, plasma ARs were analyzed by extraction with diethyl ether and normal phase liquid chromatography/tandem mass spectrometry (LC-MS/MS) after extraction with diethyl ester [30] (link) modified to a 100 mm x 2 mm column and a run time of 8 min. Matrix matched calibration was used, and two control samples were run in triplicate with each batch. Control samples and calibration curve standards were run in random order throughout each sequence. Intra-batch repeatability for each control plasma sample was < 10 % and inter-batch repeatability < 15%. Naïve and memory T lymphocyte subsets. Whole blood in EDTA tubes was stained with a mixture of APC-Cy7 mouse anti-human CD3, FITC mouse anti-human CD4, PerCP-Cy5.5 mouse anti-human CD8, PE-Cy7 mouse anti-human CD45RA and PE rat anti-human CD197 (CCR7) (BD Biosciences, Oxford, UK) in the dark for 45 min at RT. Erythrocytes were lysed using BD Pharm Lyse™ lysing buffer (BD Biosciences, Oxford, UK), incubating in the dark for 20 min at RT, before washing twice with BD CellWASH™ (BD Biosciences). Samples were analysed on a BD FACSCanto II flow cytometer. Results are reported as percentages of parent CD8 + and CD4 + T lymphocyte subsets.
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