VDE | Violaxanthin de-epoxidase
AS15 3091 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Brachypodium distachyon, Deschampsia antarctica, Oryza sativa
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Propeptide 52 kDa, mature protein: 39.7 kDa, migrates at 40 kDa (Arabidopsis thaliana)
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R – 10 µL of total protein extract of Oryza sativa (50 mg macerated leaves in 100 µL Lysis Buffer).
B – 10 µL of total protein extract of Brachypodium distachyon (50 mg macerated leaves in 100 µL Lysis Buffer).
A – 10 µL of total protein extract of Arabidopsis thaliana diluted to 1:2 (50 mg macerated leaves in 200 µL 2X Laemmli Buffer). MW marker: NZYColour Protein Marker II from NZYTech.
10 µL of total protein extracted freshly from Arabidopsis thaliana, Brachypodium distachyon, and Oryza sativa with Lysis Buffer (7M Urea, 2M Thiourea, 4% CHAPS, 35 mM Tris), and denatured in 1X Laemmli with 25 mM DTT at 95ºC for 4 min, were separated on 10 % SDS-PAGE and blotted 1h to PVDF membrane (pore size of 0.45 μm, GE Healthcare), using wet transfer. Blot was blocked with Blocking solution containing 5% milk in TBS-T, for 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1:2000 in Blocking solution containing 5% milk in TBS-T, for O/N at 4ºC, with agitation. The antibody solution was decanted, and the blot was washed 3 times for 10 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG, horse radish peroxidase conjugated) diluted to 1:20 000 in Blocking solution containing 5% milk in TBS-T, for 1h/RT with agitation. The blot was washed 6 times for 5 min in TBS-T at RT with agitation and developed for 30 seconds with Agrisera ECLSuperBright. Exposure time was 10 seconds.
Courtesy of Dr. Nelson Saibo,Plant Gene Regulation Lab (GPlantS Unit), ITQB NOVA, Portugal
Extraction buffer must contain 7 M urea for Brachypodium distachyon and Oryza sativa samples.
This product can be sold containing ProClin if requested.
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