Gssh gsh quantification kit

The 100 mg liver tissues or lens epithelial cells were obtained to be homogenized 0.5 ml 5% SSA and then centrifuged (8,000 X g, 10 min) to transfer the supernatant to a new tube and add ddH₂O to reduce the concentration of SSA from 0.5 to 1%. GSH of plasma, liver tissues, or lens epithelial cells was determined with a GSSH/GSH Quantification Kit (Dojindo; MD, U.S.A.) according to the manufacturer's instructions.

After siRNA treatment for 72 hours or K‐563 treatment for 24 hours, the Glutathione (GSH) levels in cells were measured using a GSH/GSSH‐Glo Assay Kit (Promega) or GSSH/GSH Quantification Kit (Dojindo, Kumamoto, Japan) according to the manufacturer's instructions. Either the luminescence (with the GSH/GSSH‐Glo Assay) or absorbance (with the GSSH/GSH Quantification Kit) were measured as the readout using a Top count NXT (PerkinElmer) or a SpectraMax 340PC (Molecular Devices, San Jose, CA), respectively. The GSH levels were normalized by the values observed in the negative control siRNA‐treated sample for Nrf2 siRNA or the DMSO‐treated sample for K‐563.

The liver tissue was homogenized on ice in 300 μL of malondialdehyde (MDA) lysis buffer (with 3 μL of BHT (100×) and then centrifuged (13,000 × g, 10 min) to remove insoluble material. Lipid peroxidation of the plasma was determined with a Lipid Peroxidation (MDA) Colorimetric/Fluorometric Assay Kit (BioVision; CA, U.S.A.). Glutathione peroxidase (GSH-Px) activity in plasma was measured using the Cayman (MI, U.S.) assay kit and superoxide dismutase (SOD) in the liver tissue was determined with a SOD Assay Kit-WST (Dojindo; MD, U.S.A.). GSH of liver tissues were determined with GSSH/GSH Quantification Kit (Dojindo; MD, U.S.A.) according to the manufacturer’s instructions.