GPXh | Glutathione peroxidase (algal)
AS15 2882 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Chlamydomonas reinhardtii
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18 | 15 kDa
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10 µg of a total protein from Chlamydomonas reinhardtii extracted with 2 % SDS/50 mM TRIS pH 6.8 + protease inhibitor cocktail were separated on 12 % SDS-PAGE and blotted for 1 h to PVDF using semi-dry transfer. Blots were blocked with 5 % low-fat milk powder TBS + 0.1 % Tween for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 5 000 for 1 h at RT with agitation. The antibody solution was decanted and the blot was rinsed, then washed 3 times each for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:25 000 in 2 % low-fat milk powder TBS + 0.1 % Tween for 1h at RT with agitation. The blot was washed as above and developed for 30 s with chemiluminescent detection reagent according to the manufacturer's instructions. Exposure time was typically 30 seconds.
Courtesy Dr. Thomas Roach, University of Innsbruck, Austria
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