Nrf2 transcription factor assay kit colorimetric

The Glutathione/Glutathione disulfide (GSH/GSSG) ratio was determined using the GSH/GSSG ratio detection assay kit (fluorometric - green) (#ab205811 from Abcam) and the transcriptional activity of Nrf2 was determined using the Nrf2 transcription factor assay kit (colorimetric) (#ab207223 from Abcam). Methylglyoxal (MG) formation was determined using the methylglyoxal assay kit (#ab241006 from Abcam) and 3-nitrotyrosine formation was measured using the 3-nitrotyrosine ELISA kit (#ab116691 from Abcam) according to the manufacturer’s instructions.

Antibodies recognizing NANOG (D73G4), SOX2 (D6D9), KLF4 (D1F2), HO1 (D60G11), Vimentin (D21H3) and Keap1 (D6B12) were obtained from Cell Signaling Technology (Danvers, MA, USA). E-cadherin (610404) was obtained from BD Biosciences (San Jose, CA, USA). Nrf2 (ab-89443), SLUG (ab-27568), SNAIL (ab-53519) and Bach1 (ab-115210) were purchased from Abcam, (Burlingame, CA, USA) and NQO1 (sc-32793), BAX (sc-7480), BCL2 (sc-7382) were obtained from Santa Cruz Biotechnology (Dallas, TX, USA). Nrf2-targeting shRNA lentiviral particles (sc-37030-v) were obtained from Santa Cruz Biotechnology (CA, USA). Bovine serum albumin (BSA), fibroblast growth factor (FGF) and epidermal growth factor (EGF) were purchased from Sigma-Aldrich (St. Louis, MO, USA). ALDEFLUOR Kit (01700) was obtained from STEM CELL technologies (Vancouver, Canada). Nrf2 Transcription Factor Assay Kit (Colorimetric) (ab207223) was obtained from Abcam. Annexin-V-FITC (556419) was obtained from BD Biosciences (San Jose, CA, USA).

The binding of transcription factor NF-E2 p45-related factor 2 (Nrf2) with the antioxidant response element (ARE) was assessed with an ELISA assay kit (Nrf2 Transcription Factor Assay Kit, Colorimetric, Abcam). Five micrograms of left ventricle protein was incubated with oligonucleotide ARE consensus binding site (5′ GTCACAGTGACTCAGCAGAATCTG–3′). Detection was performed with specific antibodies and measured by colorimetric (HRP catalyzed reaction) at 450 nm in a hybrid multimode reader (Sinergy H1, Biotek-Agilent, Santa Clara, CA, USA).