Toxicity of cART regimens (TDF + 3TC + EFV and AZT + 3TC + EFV) or 2-DG and DON were evaluated using AlmarBlue assay (Invitrogen) over the course of 5 days, and 48 h, respectively, according to the manufacturer’s protocol.
Almarblue assay
Manufactured by Thermo Fisher Scientific
Sourced in United States
AlmarBlue assay is a cell viability reagent that measures the metabolic activity of cells. It is a fluorometric and colorimetric assay that allows for the quantitative measurement of cell proliferation and cytotoxicity.
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Lab products found in correlation
Cellrox deep red flow cytometry assay kit, by Thermo Fisher Scientific (1 mentions)
Near ir viability, by Thermo Fisher Scientific (1 mentions)
Fortessa flow cytometer, by BD (1 mentions)
Glucose glotm assay, by Promega (1 mentions)
Glutamate glotm assay, by Promega (1 mentions)
Lactate glotm assay, by Promega (1 mentions)
3 protocols using almarblue assay
1
Evaluating Antiretroviral and Metabolic Toxicity
2
Cell Viability Assay for NCFs
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Almar Blue Assay (Invitrogen- Thermo Fischer Scientific, Carlsbad, CA, USA) was used to determine the cell viability of NCFs in the presence of the sphingolipids and TGFβ according to manufacturer’s guidelines as reported previously [14] .
3
Evaluating Cytotoxicity and Cellular Responses
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Cytotoxicity assays: Toxicity of cART regimens (TDF+3TC+EFV and AZT+3TC+EFV) or 2-DG and DON were evaluated using AlmarBlue assay (Invitrogen) over the course of five days, and 48h, respectively, according to the manufacturer's protocol.
Flow cytometry: Activation from latency was measured using GFP (J-Lat 10.6) or HIV-1 core antigen-FITC, KC57 (Beckman Coulter) staining (U1) complemented with Near-IR viability (Invitrogen). Cellular ROS was measured using CellROX TM Deep red Flow Cytometry Assay Kit (Invitrogen), according to the manufacturer's protocol. Experiments were run on Fortessa flow cytometer (BD Bioscience) and analyzed using FlowJo 10.6.2 (TreeStar Inc).
Intracellular metabolite measurement: Intracellular metabolites were measured using Glucose-Glo TM Assay, Lactate-Glo TM Assay, and Glutamate-Glo TM Assay (Promega) according to the manufacturers protocol.
Flow cytometry: Activation from latency was measured using GFP (J-Lat 10.6) or HIV-1 core antigen-FITC, KC57 (Beckman Coulter) staining (U1) complemented with Near-IR viability (Invitrogen). Cellular ROS was measured using CellROX TM Deep red Flow Cytometry Assay Kit (Invitrogen), according to the manufacturer's protocol. Experiments were run on Fortessa flow cytometer (BD Bioscience) and analyzed using FlowJo 10.6.2 (TreeStar Inc).
Intracellular metabolite measurement: Intracellular metabolites were measured using Glucose-Glo TM Assay, Lactate-Glo TM Assay, and Glutamate-Glo TM Assay (Promega) according to the manufacturers protocol.
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