Intracellular ROS generation was evaluated using the cell‐permeant Thermo Fisher CellROX Deep Red Reagent fluorogenic probe kit. HepG2 and VL‐17A cells (10 × 103 cells/well) were seeded in 96‐well plates and incubated with 50 to 100 mM EtOH, DHM, and EtOH with DHM for 24 hours. After the incubation, CellROX Reagent was added to a final concentration of 5 µM to the cells and incubated for 30 minutes at 37°C. After incubation with CellROX, medium and reagent were removed, and cells were washed 3 times with 1X PBS and measured fluorometrically using a BioTek Synergy H1 Hybrid Multi‐Mode Reader plate reader. Similar to the design of the ROS measurement assay, cytotoxicity was evaluated using the Promega Mitochondrial ToxGlo Assay Kit (Southampton, UK), a cell‐based assay that measures cytotoxicity via a fluorogenic peptide substrate (bis‐AAF‐R110), and evaluated for fluorescence according to the manufacturer’s protocol 24 hours after treatment conditions.
Synergy h1 hybrid multi mode reader plate reader
Manufactured by Agilent Technologies
Sourced in Macao
The Synergy H1 Hybrid Multi-Mode Reader is a plate reader that can perform various detection modes, including absorbance, fluorescence, and luminescence. It is designed to accommodate a range of sample types and microplate formats.
Automatically generated - may contain errors
Lab products found in correlation
Steatosis colorimetric assay kit, by Cayman Chemical (2 mentions)
Total bdnf quantikine elisa kit, by R&D Systems (2 mentions)
Alt and ast activity assay kit, by Agilent Technologies (2 mentions)
Triglyceride colorimetric assay kit, by Cayman Chemical (2 mentions)
Mitochondrial toxglo assay kit, by Promega (1 mentions)
8 protocols using synergy h1 hybrid multi mode reader plate reader
1
Intracellular ROS and Cytotoxicity Assay
2
Fluorometric Tyrosine Dimerization Assay
3
Ethanol and Acetaldehyde Metabolism Assay
4
Lipid Accumulation Quantification
5
Serum Biomarker Analysis Protocol
6
Serum Biomarker Quantification Protocol
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The activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the serum samples were measured using the Sigma ALT and AST Activity Assay Kit (St. Louis, MO) and read using the BioTek Synergy H1 Hybrid Multi‐Mode Reader plate reader (BioTek, Winooski, VT). Serum triglyceride content was measured using the Cayman Chemical Triglyceride Colorimetric Assay Kit (Cayman Chemical, Ann Arbor, MI). Serum BDNF was measured using the R&D Systems (Minneapolis, MN, USA) Total BDNF Quantikine ELISA Kit and following the kit protocol and guidelines for serum tissue analysis.
7
Intracellular Lipid Accumulation Assay
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Intracellular lipid accumulation was assessed using Cayman’s Steatosis Colorimetric Assay Kit with dye extraction, according to the manufacturer's protocol. HepG2 and VL‐17A cells (10 × 103 cells/well) were seeded in 96‐well plates and incubated with various concentrations of EtOH, DHM, and EtOH with DHM for 72 hours. Lipid accumulation was quantified using the BioTek Synergy H1 Hybrid Multi‐Mode Reader plate reader.
8
Ethanol and Acetaldehyde Production Assay
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EtOH concentration and ACH production were measured using an EtOH and ACH Enzyme Assay Kit (Megazyme, Bray, Ireland) in 96‐well plates. ACH ammonia trimer and EtOH were used as the standard according to the manufacturer’s instructions for the ACH assay and EtOH assay, respectively. VL‐17A cells and HepG2 cells were incubated with 50 mM EtOH and vehicle (0.2% DMSO) or 100 nM to 50 µM DHM, or untreated for 2 hours before measurements using the BioTek Synergy H1 Hybrid Multi‐Mode Reader plate reader (BioTek, Winooski, VT).
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