Protein carbonyl colorimetric assay kit

A375 cells were plated in 100 mm dishes, cultured overnight and next day were treated with L-SK-4 (100 μM). After trypsinization, pellets were collected, resuspended and sonicated before the Protein Carbonyl Colorimetric Assay Kit (Cambridge Bioscience Ltd., Cambridge, UK) was utilized for the determination of protein carbonyl content according to the manufacture’s protocol.

A375 cells were plated in 100 mm dishes (1.4 × 10⁶, 0.7 × 10⁶ and 0.4 × 10⁶ per dish for 24, 48 and 72 h, respectively) and cultured overnight. The next day, cells were treated with either N-acetyl cysteine (NAC) (2.5 mM), tert-butyl hydroperoxide (TBH) (200 μΜ), synthetic PEITC (28, 12 and 7 μM) or edible (16, 10 and 9 μM) or non-edible (32, 18 and 13 μM) watercress samples for 24, 48 and 72 h, respectively. After trypsinization, pellets were collected, re-suspended in PBS and sonicated. For the determination of lipid peroxidation content, the whole suspension was further diluted with 4 mL of 4% v/v acetic acid solution containing 8% TBA and 0.1% SDS. The final mixture was heated at 95 °C for 1 h and centrifuged at 3000 rpm for 2 min. The TBARS Assay kit (Cambridge Bioscience Ltd., Cambridge, UK) was utilized for the determination of malondialdehyde (MDA) content according to the manufacture’s protocol. For the determination of protein carbonyl content, cells were trypsinized and pellets were collected, re-suspended in PBS (supplemented with 1 mM EDTA) and sonicated. The Protein Carbonyl Colorimetric Assay Kit (Cambridge Bioscience Ltd., UK) was used according to the manufacture’s protocol.