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Aqua live dead amine dye amcyan

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The Aqua Live/Dead amine dye-AmCyan is a fluorescent dye used to distinguish between live and dead cells in a sample. It binds to the amines present in cells, providing a clear visualization of cell viability.

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Lab products found in correlation

Anti cd8 qdot705 clone 3b5, by Thermo Fisher Scientific (4 mentions) Anti cd28 ecd clone cd28.2, by Beckman Coulter (3 mentions) Anti cxcr3 alexafluor 488 clone 1c6 cxcr3, by BD (2 mentions) Anti cd161 pe clone hp 3g10 and anti il 17 alexa488 clone ebio64dec17, by Thermo Fisher Scientific (2 mentions) Anti cd161 bv421 clone hp 3g10, by BioLegend (2 mentions) Annexin 5 buffer, by BD (2 mentions) Ifn γ bv510, by BioLegend (1 mentions) Cd4 bv605, by BD (1 mentions) Tnf α alexafluor 700, by BD (1 mentions) Cd69 pe cf594, by BD (1 mentions) Cd161 pe, by BioLegend (1 mentions) Cd8 bv650, by BD (1 mentions) Fortessa, by BD (1 mentions) Cd3 apc cy7, by BD (1 mentions) Prism version 6, by GraphPad (1 mentions) Cytofix cytoperm, by BD (1 mentions)

7 protocols using aqua live dead amine dye amcyan

1

Multiparametric Flow Cytometry Analysis

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Fourteen-parameter flow cytometric analysis was performed on PB-, LN- and RB-derived cells. Predetermined optimal concentrations were used of the following antibodies: anti-CD3-APC-Cy7 (clone SP34-2), anti-Ki-67-Alexa700 (clone B56), anti-IFN-γ-PE-Cy7 (clone B27), anti-CD8-PE-CF-594 (clone RPA-T8), anti-TNFα-Alexa700 (clone MAb11), (all from BD Pharmingen); anti-IL-17-Alexa Fluor488 (clone eBio64DEC17), anti-IL-22-APC (clone IL22JOP) (all from eBioscience); anti-CD4-BV421 (clone OKT4), anti-IL-2-BV605 (clone MQ1-17H12), (all from Biolegend); anti-CD8-Qdot705 (clone 3B5) and Aqua Live/Dead amine dye-AmCyan (all from Invitrogen). Flow cytometric acquisition was performed on at least 100,000 CD3+ T cells on an LSRII cytometer driven by the FACS DiVa software. Analysis of the acquired data was performed using FlowJo software (TreeStar).
Ryan E.S., Micci L., Fromentin R., Paganini S., McGary C.S., Easley K., Chomont N, & Paiardini M. (2016). Loss of Function of Intestinal IL-17 and IL-22 Producing Cells Contributes to Inflammation and Viral Persistence in SIV-Infected Rhesus Macaques. PLoS Pathogens, 12(2), e1005412.
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2

Comprehensive Immune Profiling by Flow Cytometry

Cited 1 time
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Fourteen-parameter flow cytometric analysis was performed on peripheral blood, LN, and RB derived cells according to standard procedures using a panel of monoclonal antibodies that we and others have shown to be cross-reactive with SMs and RMs.12 (link) The following antibodies were used at predetermined optimal concentrations: anti-CCR6-PE-Cy7 (clone 11A9), anti-CCR5-PE and -APC (clone 3A9), anti-CD3-APC-Cy7 (clone SP34-2), anti-CD62L-FITC (clone SK11), anti-CD95-PE-Cy5 (clone DX2), anti-Ki-67-Alexa700 (clone B56), anti-IFN-γ-Alexa700 (clone B27), and anti-CXCR3-AlexaFluor 488 (clone 1C6/CXCR3) from BD Biosciences; anti-CD161-PE (clone HP-3G10) and anti-IL-17-Alexa488 (clone eBio64DEC17) from eBioscience; anti-CD28-ECD (clone CD28.2) from Beckman Coulter; anti-CD4-BV421 (clone OKT4), anti-CD4-BV605 (clone OKT4), and anti-CD161-BV421 (clone HP-3G10) from Biolegend; anti-CD8-Qdot705 (clone 3B5) and Aqua Live/Dead amine dye-AmCyan from Invitrogen. Flow cytometric acquisition was performed on at least 100,000 CD3+ T cells on an LSRII cytometer driven by the FACS DiVa software, or at least 10,000 CD3+ T cells for rectal biopsy-derived cells. The data acquired were analyzed using FlowJo software (version 9.8.5; TreeStar).
McGary C.S., Alvarez X., Harrington S., Cervasi B., Ryan E.S., Iriele R.I., Paganini S., Harper J., Easley K., Silvestri G., Ansari A.A., Lichterfeld M., Micci L, & Paiardini M. (2017). The loss of CCR6+ and CD161+ CD4+ T cell homeostasis contributes to disease progression in SIV-infected rhesus macaques. Mucosal immunology, 10(4), 1082-1096.
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3

Multicolor Flow Cytometry for Immune Profiling

Cited 1 time
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Multi-color flowcytometric analysis was performed on cells according to standard procedures using anti-human mAbs that cross-react with rhesus macaques. The following antibodies were used at predetermined optimal concentrations: CD45 (BD clone D058-1283), CD3 APC-Cy7 (BD clone SP34-2), CD4 BV605 (BD clone L200), CD8 BV650 (BD clone SK1), CD69 PE-CF594 (BD clone FN50), CD161 PE (Biolegend clone HP- 3G10), TCR γδ PE-Cy7 (BD clone B1), TCR Vδ1 FITC (ThermoScientific clone TS8.2), TCR Vδ2 FITC (ThermoScientific clone 15D), TCR Vα7.2 BV421 (Biolegend clone 3C10), and Aqua Live/Dead amine dye-AmCyan from Invitrogen (Waltham, MA). Surface staining was carried out by standard procedures as earlier described (23 (link)). Flow cytometric acquisition was performed on the BD Fortessa instrument driven by the FACS DiVa software for at least 100,000 CD3+ T cells in PBMC or at least 10,000 CD3+ T cells for rectal biopsy lymphocytes. The data acquired were analyzed using FlowJo software (version 10.7.1; TreeStar, Ashland, OR). For evaluation of cytokine production, intracellular cytokine staining with IFN-γ BV510 (Biolegend clone 4S.B3), IL-17 PerCP-Cy5.5 (eBioscience clone eBio64DEC17), IL-22 APC (Invitrogen clone IL22JOP), and TNF-α Alexa Fluor 700 (BD clone MAb11) were utilized.
Walker E.M., Slisarenko N., Gerrets G.L., Grasperge B.F., Mattison J.A., Kissinger P.J., Welsh D.A., Veazey R.S., Jazwinski S.M, & Rout N. (2021). Dysregulation of IL-17/IL-22 Effector Functions in Blood and Gut Mucosal Gamma Delta T Cells Correlates With Increase in Circulating Leaky Gut and Inflammatory Markers During cART-Treated Chronic SIV Infection in Macaques. Frontiers in Immunology, 12, 647398.
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4

Annexin V Staining of PBMCs

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Freshly isolated PBMCs from 8 healthy RMs (2×106 cells) were washed once with Annexin V buffer (BD Biosciences). PBMCs were then stained ex vivo with surface markers for CD3, CD4, CD8, CCR6, CD161, CD28, and CD95, as well as with Annexin V (APC; BD Biosciences) and Aqua Live/Dead amine dye-AmCyan (Invitrogen) for 30 min at room temperature. Following staining, cells were washed with Annexin V buffer and immediately acquired on an LSRII cytometer. 4 × 106 freshly isolated PBMCs from the same animals were also cultured for 24 hours in complete RPMI media prior to performing Annexin V staining, as described above.
McGary C.S., Alvarez X., Harrington S., Cervasi B., Ryan E.S., Iriele R.I., Paganini S., Harper J., Easley K., Silvestri G., Ansari A.A., Lichterfeld M., Micci L, & Paiardini M. (2017). The loss of CCR6+ and CD161+ CD4+ T cell homeostasis contributes to disease progression in SIV-infected rhesus macaques. Mucosal immunology, 10(4), 1082-1096.
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5

Comprehensive Immune Profiling by Flow Cytometry

Cited 1 time
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Fourteen-parameter flow cytometric analysis was performed on peripheral blood, LN, and RB derived cells according to standard procedures using a panel of monoclonal antibodies that we and others have shown to be cross-reactive with SMs and RMs.12 (link) The following antibodies were used at predetermined optimal concentrations: anti-CCR6-PE-Cy7 (clone 11A9), anti-CCR5-PE and -APC (clone 3A9), anti-CD3-APC-Cy7 (clone SP34-2), anti-CD62L-FITC (clone SK11), anti-CD95-PE-Cy5 (clone DX2), anti-Ki-67-Alexa700 (clone B56), anti-IFN-γ-Alexa700 (clone B27), and anti-CXCR3-AlexaFluor 488 (clone 1C6/CXCR3) from BD Biosciences; anti-CD161-PE (clone HP-3G10) and anti-IL-17-Alexa488 (clone eBio64DEC17) from eBioscience; anti-CD28-ECD (clone CD28.2) from Beckman Coulter; anti-CD4-BV421 (clone OKT4), anti-CD4-BV605 (clone OKT4), and anti-CD161-BV421 (clone HP-3G10) from Biolegend; anti-CD8-Qdot705 (clone 3B5) and Aqua Live/Dead amine dye-AmCyan from Invitrogen. Flow cytometric acquisition was performed on at least 100,000 CD3+ T cells on an LSRII cytometer driven by the FACS DiVa software, or at least 10,000 CD3+ T cells for rectal biopsy-derived cells. The data acquired were analyzed using FlowJo software (version 9.8.5; TreeStar).
McGary C.S., Alvarez X., Harrington S., Cervasi B., Ryan E.S., Iriele R.I., Paganini S., Harper J., Easley K., Silvestri G., Ansari A.A., Lichterfeld M., Micci L, & Paiardini M. (2017). The loss of CCR6+ and CD161+ CD4+ T cell homeostasis contributes to disease progression in SIV-infected rhesus macaques. Mucosal immunology, 10(4), 1082-1096.
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6

Twelve-parameter Flow Cytometry Analysis

Cited 2 times
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Twelve-parameter flow cytometric analysis was performed on WB, LN, RB and BM derived cells according to standard procedures using a panel of monoclonal antibodies that we and others have shown to be cross-reactive with RM [35] (link)–[37] (link). Predetermined optimal concentrations were used of the following antibodies: anti-CD3-Alexa700 (clone SP34-2), anti-CD3-APC-Cy7 (clone SP34-2), anti-CD4-Pacific Blue (clone OKT4), anti-CD8-APC-Cy7 (clone SK1), anti-CD95-PE-Cy5 (clone DX2), anti-CCR5-APC (clone 3A9), anti-Ki-67-Alexa700 (clone B56), anti-Ki-67-FITC (clone B56), anti-CD14-Pe-Cy7 (clone M5E2), anti-CD16-BV421 (clone 3G8), anti-CD62L-PE (clone SK11), anti-CCR7-PE-Cy7 (Clone 3D12) (all from BD Pharmingen); anti-CD28-ECD (clone CD28.2) (Beckman Coulter); anti-CD8-Qdot705 (clone 3B5) and Aqua Live/Dead amine dye-AmCyan (Invitrogen). Intracellular staining for Ki-67 was performed at room temperature for 30 minutes following permeabilization with cytofix/cytoperm (BD Bioscience). Flow cytometric acquisition was performed on an LSRII cytometer driven by the FACS DiVa software. Analysis of the acquired data was performed using FlowJo software (TreeStar) and graphs were prepared using Prism version 6.0 (GraphPad).
Micci L., Alvarez X., Iriele R.I., Ortiz A.M., Ryan E.S., McGary C.S., Deleage C., McAtee B.B., He T., Apetrei C., Easley K., Pahwa S., Collman R.G., Derdeyn C.A., Davenport M.P., Estes J.D., Silvestri G., Lackner A.A, & Paiardini M. (2014). CD4 Depletion in SIV-Infected Macaques Results in Macrophage and Microglia Infection with Rapid Turnover of Infected Cells. PLoS Pathogens, 10(10), e1004467.
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7

Annexin V Staining of PBMCs

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Freshly isolated PBMCs from 8 healthy RMs (2×106 cells) were washed once with Annexin V buffer (BD Biosciences). PBMCs were then stained ex vivo with surface markers for CD3, CD4, CD8, CCR6, CD161, CD28, and CD95, as well as with Annexin V (APC; BD Biosciences) and Aqua Live/Dead amine dye-AmCyan (Invitrogen) for 30 min at room temperature. Following staining, cells were washed with Annexin V buffer and immediately acquired on an LSRII cytometer. 4 × 106 freshly isolated PBMCs from the same animals were also cultured for 24 hours in complete RPMI media prior to performing Annexin V staining, as described above.
McGary C.S., Alvarez X., Harrington S., Cervasi B., Ryan E.S., Iriele R.I., Paganini S., Harper J., Easley K., Silvestri G., Ansari A.A., Lichterfeld M., Micci L, & Paiardini M. (2017). The loss of CCR6+ and CD161+ CD4+ T cell homeostasis contributes to disease progression in SIV-infected rhesus macaques. Mucosal immunology, 10(4), 1082-1096.
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