AS21 4532 Anti-AOX2 | Plant alternative oxidase 2

Samples:
Total protein 1-4
Cytoplasmic fraction
5-8 Nuclear fraction 9-12

20 µg/well of total protein extracted freshly from fractionation of Arabidopsis thaliana 7-day-old green seedlings and denatured with 2x Laemmli Sample Buffer at 90°C for 5 min were separated on 12 % SDS-PAGE  and blotted 1h to nitrocellulose (pore size of  0.45  um), using wet transfer. Blot was blocked with 5% milk for 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for ON/4°C with agitation in TBS-T. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:25 000 in  for 1h/RT with agitation. The blot was washed as above and developed for 5  min with Agrisera ECLBright. Exposure time was 200 seconds.

Courtesy of Dr. Zhengyao Shao, The University of Texas at Austin , USA

Alternative oxidases (AOX) are quinol oxidases located in the inner mitochondrial membrane of plants. They function as terminal oxidases in the alternate electron transport pathway, oxidizing ubiquinone to reduce oxygen to water.

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