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Fixable viability dye

Manufactured by Cell Signaling Technology
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Fixable Viability Dye is a fluorescent reagent that can be used to distinguish between live and dead cells. It binds to cellular proteins, allowing for the identification of non-viable cells in flow cytometry and other applications.

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Lab products found in correlation

Rorγt, by Cell Signaling Technology (2 mentions) Ifn γ, by Cell Signaling Technology (1 mentions) Malt1, by Cell Signaling Technology (1 mentions) Gm csf, by Cell Signaling Technology (1 mentions) Il 17a, by Cell Signaling Technology (1 mentions) Foxp3, by Cell Signaling Technology (1 mentions) T bet, by Cell Signaling Technology (1 mentions) Il 17a, by Thermo Fisher Scientific (1 mentions) Thy 1, by Thermo Fisher Scientific (1 mentions) Foxp3 transcription factor staining kit, by Thermo Fisher Scientific (1 mentions) γδtcr, by BioLegend (1 mentions) C maf, by Cell Signaling Technology (1 mentions) P drp1, by Cell Signaling Technology (1 mentions) Ifn γ, by Thermo Fisher Scientific (1 mentions)

2 protocols using fixable viability dye

1

Comprehensive Immune Cell Profiling

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Cells were stained with the following antibodies: 0.1 µl/100 μl Fixable Viability Dye (eFluor 520, eFluor 780), 0.25 µg/100 μl CD4 (Brilliant violet 711, clone: GK1.5), 0.5 µg/100 μl RORγt (APC, clone: AFKJS-9), 0.4 µg/100 μl T-bet (PE-cyanine7, clone: 4B10), 0.5 µg/100 μl IL-17A (eFluor 450, clone: eBio17B7), 0.25 µg/100 μl GM-CSF (PE, clone: MP1-22E9), 0.25 µg/100 μl IFNγ (FITC, clone: XMG1.2), 0.5 µg/100 μl CD25 (PE-Cy7, clone: PC61), 0.06 µg/100 μl Foxp3 (eFluor 450, FITC, clone: FJK-16s), 0.5 µg/100 μl CD8α (PE, clone: 53-6.7), 5 µg/100 μl IL-23R (PE, BV711, clone: O78-1208), 0.5 µg/100 μl CCR6 (BV605, clone: 29-2L17), 1 µg/100 μl CD29 (PE-Cy7, clone: HMβ1-1), 2 µl/100 μl Malt1 (#2494 Cell Signaling Technology, Danvers, MA, USA), 5 µg/ml Alexa Fluor 488 goat anti-rabbit IgG (H + L) (A11008, Fisher Scientific, Thermo Fisher Scientific, PA, USA), 2 µl/100 μl pStat3 Y705 (#4324, Alexa Fluor 647, clone: D3A7), 0.25 µg/100 μl CD44 (APC, clone: IM7), 0.5 µg/100 μl CD62L (PE, clone: MEL-14), 0.5 µg/100 μl CD69 (PerCP-Cy5.5, clone: H1.2F3), 0.25 µg/100 μl Ki-67 (FITC, clone: SolA15), 1 µl/100 μl NF-κB p65 (#4764, clone: C22B4), 1 µl/100 μl RelB (#4922, clone: C1E4), 5 µg/ml GFP (A21311, Alexa Fluor 488), 0.125 µg/100 μl CD90.1 (PE-Cy7, clone: HIS51).
Cho J.J., Xu Z., Parthasarathy U., Drashansky T.T., Helm E.Y., Zuniga A.N., Lorentsen K.J., Mansouri S., Cho J.Y., Edelmann M.J., Duong D.M., Gehring T., Seeholzer T., Krappmann D., Uddin M.N., Califano D., Wang R.L., Jin L., Li H., Lv D., Zhou D., Zhou L, & Avram D. (2019). Hectd3 promotes pathogenic Th17 lineage through Stat3 activation and Malt1 signaling in neuroinflammation. Nature Communications, 10, 701.
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2

Multiparametric Flow Cytometry Analysis

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Fluorescent antibodies were purchased from Biolegend (CD3, γδTCR, CD4, CD8, CD27, Vγ4, Vγ5, IL-17A, IFN-γ, CD25, CD73, Thy-1.2, Anti-rat IgM), eBioscience (Fixable Viability Dye, RORγt, c-Maf, CD44, CD24), and Cell Signaling Technologies (p-Drp1). For cell surface marker staining, cells were first treated with Fc Blocker and then stained with cell surface antibodies at 4°C for 20 min. The relevant isotype control antibodies were also used. For intracellular cytokine staining, cells were stained with surface antibodies followed by fixation, permeabilization (Biolegend), and finally intracellular staining of IL-17/IFN-γ at 4°C for 2h. For c-Maf/RORγt staining, cells were stained with surface antibodies (Abs), and then fixed and permeabilized with Foxp3/Transcription Factor Staining kit (eBioscience) for 45 min followed by intranuclear staining of c-Maf/RORγt at 4°C for 2h.
Wang Y., Qin H., Cai Y., Chen X., Li H., Montoya-Durango D.E., Ding C., Hu X., Chariker J.H., Sarojini H., Chien S., Rouchka E.C., Zhang H.G., Zheng J., Qiu F, & Yan J. (2023). Natural γδT17 cell development and functional acquisition is governed by the mTORC2-c-Maf-controlled mitochondrial fission pathway. iScience, 26(5), 106630.
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