Ccr5 pe dazzle 594

Manufactured by BioLegend

CCR5 PE-Dazzle 594 is a fluorescently-labeled antibody that binds to the CCR5 chemokine receptor expressed on the surface of certain immune cells. It is designed for use in flow cytometry applications to identify and quantify CCR5-positive cell populations.

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2 protocols using ccr5 pe dazzle 594

Multiparameter Flow Cytometry Analysis

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The following fluorochrome-conjugated anti-human mAb were used for flow cytometry studies: LAG3 BV650, TIGIT PE-Cy7, CTLA4 PE, CD25 PE-Dazzle 594, TIM3 PerCP-Cy5.5, CD19 Alexa Fluor 700, CCR4 BV421, ICOS PE-Cy7, CCR5 PE-Dazzle 594, HLA-DR PE, CCR7 FITC, CD38 BV711, PD-L1 BV711, CCR6 Alexa Fluor 700, PD-1 BV421, CD40L BV605, perforin PE-Dazzle 594, and CD8 PerCP from BioLegend (San Diego, CA); CD3 BUV496, CD4 APC-Cy7, CD4 APC-H7, CD127 BV605, Ki-67 PerCP-Cy5.5, PD-1 BV650, CXCR3 BV605, CD69 BV650, IL-2 BV711, CXCR5 Alexa Fluor 647, IFN-γ PE-Cy7, TNF-α FITC, granzyme B Alexa Fluor 700 and CD27 BV480 from BD Biosciences (San Jose, CA); IL-21 PE from eBioscience, (San Diego, CA); and CD45RO PE-Cy5.5 from Beckman Coulter (Fullerton, CA). LIVE/DEAD Fixable Blue Dead Cell Stain Kit from Thermo Fisher Scientific (Boston, MA) was used to detect and exclude dead cells. All the reagents were tested and titrated for optimum concentration before usage.

Rinaldi S., de Armas L., Dominguez-Rodríguez S., Pallikkuth S., Dinh V., Pan L., Gӓrtner K., Pahwa R., Cotugno N., Rojo P., Nastouli E., Klein N., Foster C., De Rossi A., Giaquinto C., Rossi P., Palma P, & Pahwa S. (2021). T cell immune discriminants of HIV reservoir size in a pediatric cohort of perinatally infected individuals. PLoS Pathogens, 17(4), e1009533.

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Multiparametric Flow Cytometry of Immune Cells

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The following fluorochrome-conjugated antibodies were used in this study: CD14 BV510, CD19 BV510, CD3 APC Fire 750 or CD3 BV605, CD4 PE/Dazzle 594, or CD4 APC Fire 750, CD8 BV421, CCR5 PE/Dazzle 594, CD56 PeCy7 (Biolegend) for surface antigens; and IFN-γ PeCy7, TNF-α FITC (Biolegend) and IL-2 PercP eFluor710 (eBioscience), for intracellular staining. PBMC were washed in PBS, and surface stained at 4°C for 20 min with saturating concentrations of different combinations of antibodies in the presence of fixable live/dead stain (Invitrogen). Cells were then fixed and permeabilized for detection of intracellular antigens. Cells were acquired on a BD Fortessa X20 using BD FACSDiva8.0 (BD Bioscience) and analysed using FlowJo 10 (Tree Star). Stochastic neighbor embedding (SNE) analysis was performed using the mrc.cytobank platform.

Gupta R.K., Abdul-jawad S., McCoy L.E., Mok H.P., Peppa D., Salgado M., Martinez-Picado J., Nijhuis M., Wensing A.M., Lee H., Grant P., Nastouli E., Lambert J., Pace M., Salasc F., Monit C., Innes A., Muir L., Waters L., Frater J., Lever A.M., Edwards S., Gabriel I.H, & Olavarria E. (2019). HIV-1 remission following CCR5Δ32/Δ32 haematopoietic stem cell transplantation. Nature, 568(7751), 244-248.

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