Qdot 585

Manufactured by Thermo Fisher Scientific

The Qdot 585 is a fluorescent nanoparticle product offered by Thermo Fisher Scientific. It is designed to emit light at a wavelength of 585 nanometers when excited by a light source.

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Lab products found in correlation

Foxp3 transcription factor staining kit, by Thermo Fisher Scientific (2 mentions) Lsrfortessa, by BD (2 mentions) Live dead aqua, by Thermo Fisher Scientific (2 mentions) Qdot 605, by Thermo Fisher Scientific (1 mentions) Brilliant violet 421, by BioLegend (1 mentions) Brilliant violet 785, by BioLegend (1 mentions) Horizon v500, by BD (1 mentions) Brilliant violet 711, by BioLegend (1 mentions) Lsrii instrument, by BD (1 mentions) Streptavidin qdot605, by Thermo Fisher Scientific (1 mentions) Facsymphony a5, by BD (1 mentions) Nkg2c, by R&D Systems (1 mentions)

3 protocols using qdot 585

Multiparametric Flow Cytometry for Immune Phenotyping

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Antibodies and clones against the following proteins were used: CD3 (UCHT1, PE-Cy5, Beckman Coulter), CD14 (MϕP9, Horizon V500, BD Biosciences), CD16 (3G8, Brilliant Violet 711 or Brilliant Violet 785, Biolegend), CD19 (HIB19, Horizon V500, BD Biosciences), CD38 (HIT2, Brilliant Violet 711, BD Biosciences), CD45 (HI30, Alexa Fluor 700, Biolegend), CD49a (TS2A, AlexaFluor 647, Biolegend), CD56 (N901, ECD, Beckman Coulter, or HCD56, Brilliant Violet 711, Biolegend), NKG2A (Z1991.10, APC-A780, Beckman Coulter), CD69 (TP1.55.3, ECD, Beckman Coulter), CXCR3 (G025H7, PE-Cy7, Biolegend). After washing twice, cells were stained with streptavidin Qdot 605 or Qdot 585 (both Invitrogen) and Live/Dead Aqua (Molecular probes, Life Technologies). After surface staining, PBMC were fixed and permeabilized using FoxP3/Transcription Factor staining kit (eBioscience). For intracellular staining, the following antibodies were used: granzyme B (GB-11, PE-CF594, BD Biosciences), IFN-γ-Brilliant Violet 570 (4S.B3, Brilliant Violet 570, Biolegend), Ki67 (B56, A700, BD Biosciences), perforin (dG9, PE, Biolegend), and TNF (MAb11, Brilliant Violet 421, Biolegend). IAV infection was monitored using anti-influenza A nucleoprotein-1 (431, Abcam). Samples were analyzed on a BD LSRFortessa equipped with 5 lasers (BD Biosciences), and data were analyzed using FlowJo versions 9.5.2 and 10.5.3 (Tree Star Inc).

Scharenberg M., Vangeti S., Kekäläinen E., Bergman P., Al-Ameri M., Johansson N., Sondén K., Falck-Jones S., Färnert A., Ljunggren H.G., Michaëlsson J., Smed-Sörensen A, & Marquardt N. (2019). Influenza A Virus Infection Induces Hyperresponsiveness in Human Lung Tissue-Resident and Peripheral Blood NK Cells. Frontiers in Immunology, 10, 1116.

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Comprehensive T Cell Profiling in HIV Infection

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Cells were stained by standard protocols as described previously.^{[15 (link),16 (link)]} The following antibodies were used to define T cell subsets (CD3, CD4, CD8, γδTCR), T cell activation (CD38, HLA-DR, CCR5, CD69, PD-1), and T cell differentiation into naïve and memory T cells, including tissue resident memory cells, (CCR7, CD45RA, CD69). The analysis of γδ T cells was included because γδ T cells are frequently found at mucosal surfaces, are rapidly depleted in HIV infection, and have also been demonstrated to serve as a viral reservoir for HIV.^{[17 (link),18 (link)]} All antibodies were obtained from BD Biosciences (San Jose, CA), except for PD-1 that was obtained from eBioscience (San Diego, CA). A live/dead cell discriminator dye (Qdot 585; Invitrogen, ThermoFisher Scientific; Waltham, MA) was included in all samples. Samples were acquired on a LSRII instrument (Beckton Dickinson, San Jose, CA) recording a minimum of 300,000 events. Samples were analyzed with FlowJo Software (Treestar, Ashland, OR) applying Boolean gating strategies.

Weber M.D., Andrews E., Prince H.A., Sykes C., Rosen E.P., Bay C., Shaheen N.J., Madanick R.D., Dellon E.S., Paris K.D., Nelson J.A., Gay C.L, & Kashuba A.D. (2018). Virologic and Immunologic Responses to Raltegravir and Dolutegravir in the Gut-Associated Lymphoid Tissue of HIV-Infected Men and Women. Antiviral therapy, 23(6), 495-504.

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Multiparametric Flow Cytometry Analysis

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Antibodies and clones used for phenotyping, intracellular staining, and cell sorting are listed in SI Appendix, Table S2. Secondary staining was performed with streptavidin Qdot 605 or Qdot 585 (both Invitrogen), anti-mouse IgM (II/41, eFluor 650NC; eBioscience), or streptavidin BB-630 or streptavidin-BV650 (BD Biosciences) and Live/Dead Aqua (Invitrogen). After surface staining, cells were fixed and permeabilized using a FoxP3/Transcription Factor staining kit (eBioscience) for subsequent intracellular staining. Purified NKG2C (no. 134591; R&D Systems) was biotinylated using a FluoReporter Mini-Biotin XX protein labeling kit (Life Technologies) and detected using streptavidin-Qdot 585, 605, or 655 (Invitrogen).
Samples were analyzed on a BD LSRFortessa equipped with four lasers (BD Biosciences) or a BD FACSymphony A5 equipped with five lasers (BD Biosciences), and data were analyzed using FlowJo version 9.5.2 and version 10.6.1 (Tree Star). UMAPs were constructed in FlowJo 10.6.1 using the UMAP plugin. UMAP coordinates and protein expression data were subsequently exported from FlowJo, and protein expression for each parameter was normalized to a value between 0 and 100. UMAP plots were made in R using ggplot, and color scale shows log2(normalized protein expression +1).

Brownlie D., Scharenberg M., Mold J.E., Hård J., Kekäläinen E., Buggert M., Nguyen S., Wilson J.N., Al-Ameri M., Ljunggren H.G., Marquardt N, & Michaëlsson J. (2021). Expansions of adaptive-like NK cells with a tissue-resident phenotype in human lung and blood. Proceedings of the National Academy of Sciences of the United States of America, 118(11), e2016580118.

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