Cd41a percp cy5

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CD41a-PerCP/Cy5.5 is a fluorescently-labeled antibody reagent used for the detection and quantification of CD41a, also known as the platelet glycoprotein IIb (GPIIb) subunit, on the surface of cells. The antibody is conjugated to the PerCP/Cy5.5 fluorochrome, which allows for multicolor flow cytometric analysis.

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PerCP-Cy5.5 (133 citations) CD41a (2 citations)

2 protocols using cd41a percp cy5

Comprehensive EV Characterization and Cytokine Profiling

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To characterize EVs, platelet-free plasma samples were stained and acquired using 14 different fluorochrome-conjugated antibodies in three separate panels, including CD235a-FITC, CD62p-APC, CD3-PerCP/Cy5.5, CD19-A700, CD28-FITC, CD62L-APC, CD11b-PE/Cy7, CD154-APC, CD41a-PerCP/Cy5.5, CD66b-PE (Biolegend), CD15-FITC (ExAlpha), CD108a-PE, CD16-V450, and CD142-PE (BD Biosciences). Samples were washed with a 0.22-μm centrifugal filter (Millipore) at 850× g for 3 minutes. EVs were then harvested from the top of the filter after washing, and data were acquired on a flow cytometer ((BD LSR II, BD Biosciences). In addition, 27 cytokines were measured using human cytokine chemokine kits (Multiplex MAG kits, Millipore): granulocyte-macrophage colony-stimulating factor (GM-CSF), IFN-γ, IL-10, IL-12(p70), IL-17A, IL-1β, IL-2, IL-21, IL-23, IL-6, IL-7, IL-8, CXCL11/ITAC, CCL3/MIP-1α, CCL4/MIP-1β, tumor necrosis factor-α, CXCL10/IP-10, epidermal growth factor (EGF), fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF), β2 microglobulin, cystatin C, myeloperoxidase, platelet-derived growth factor-AB/BB, CCL5/RANTES, soluble intercellular adhesion molecule-1, and soluble vascular cell adhesion molecule-1. A suspension array instrument (Bio-Plex 200, Bio-Rad) was used for cytokine data acquisition.

Spinella P.C., Sniecinski R.M., Trachtenberg F., Inglis H.C., Ranganathan G., Heitman J.W., Szlam F., Danesh A., Stone M., Keating S.M., Levy J.H., Assmann S.F., Steiner M.E., Doctor A, & Norris P.J. (2019). Effects of blood storage age on immune, coagulation, and nitric oxide parameters in transfused patients undergoing cardiac surgery. Transfusion, 59(4), 1209-1222.

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Flow Cytometric Analysis of Extracellular Vesicles

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Extracellular vesicles phenotyping was performed using a flow cytometer (FC) as previously described [15, 16]. Briefly, 20 μl of the supernatant of each RBC product was stained with the following linage‐specific monoclonal antibodies to identify the cell of origin of EVs: CD41a‐PerCP‐Cy5.5, CD142‐APC, CD66b‐PE, CD144‐BV421, CD235a‐FITC, CD3‐FITC, and CD14‐PE‐Cy7 (Biolegend, San Diego, CA, USA), and CD16‐ECD, CD19‐PerCP‐Cy5.5 and CD62P‐APC (BD Biosciences, Chatham, NJ, USA). Stained samples were run through an LSRII flow cytometer (BD Biosciences); sufficient events were collected to provide approximately ≥5·000 gated EV events. An AbC Anti‐Mouse Bead Kit (Life Technologies, Carslbad, CA, USA) was used to set the compensation along with the single‐stained compensation control. Small size beads, ranging from 0.2 to 1 μm (Megmix‐Plus SSC beads, Biocytex, Marseille, France), were used to generate the EV gate and to further classify them based on their size (only EVs ≤ 1.0 µm in diameter were analysed). BD TruCOUNT tubes (BD Biosciences) were used to obtain the absolute number of EVs/μL. Data were analysed using FlowJo v10.

Wirtz M.R., Almizraq R.J., Weber N.C., Norris P.J., Pandey S., Spinella P.C., Muszynski J.A., P. Acker J, & Juffermans N.P. (2020). Red‐blood‐cell manufacturing methods and storage solutions differentially induce pulmonary cell activation. Vox Sanguinis, 115(5), 395-404.

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