Detection of Ascorbate Peroxidase (APX) Activity in Plant Tissues: Using Non-denaturing PAGE and Spectrophotometric Assay.
Marta Rodríguez-RuizHayet HoumaniMaría A Muñoz-VargasJosé M PalmaFrancisco J CorpasPublished in: Methods in molecular biology (Clifton, N.J.) (2024)
At the cellular level, the generation of reactive oxygen species (ROS), such as hydrogen peroxide (H 2 O 2 ), due to different abiotic or biotic stress, causes oxidative stress that induces an imbalance in the metabolism. Among the different H 2 O 2 -scavenging enzymatic antioxidants, ascorbate peroxidase (APX) is a heme-peroxidase that plays an important role in the ascorbate-glutathione pathway using ascorbate to reduce H 2 O 2 to water. Using non-denaturing polyacrylamide gel electrophoresis (PAGE) in combination with a spectrophotometric assay for APX activity, the protocol allows identifying diverse APX isozymes present in different organs and plant species.
Keyphrases
- hydrogen peroxide
- reactive oxygen species
- nitric oxide
- oxidative stress
- high throughput
- dna damage
- simultaneous determination
- gene expression
- randomized controlled trial
- cell death
- mass spectrometry
- real time pcr
- induced apoptosis
- high resolution
- diabetic rats
- hyaluronic acid
- label free
- endoplasmic reticulum stress
- liquid chromatography