PsbY | Small subunit Y of PSII
AS06 114 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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1 µg of total protein from Arabidopsis thaliana PSII enriched membranes in BBY storage buffer (20 mM MES-NaOH pH 6.3, 400 mM sorbitol, 5 mM MgCl2, 10 mM CaCl2, and 15 mM NaCl) denatured with Laemmli sample buffer at 70C for 5 min were separated on 16,5 % Tris-Tricine, 6 M Urea SDS-PAGE with a 10 % spacer gel, and blotted 1h to PVDF using semi-dry transfer. Blots were blocked with 2 % low fat milk for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for 1h at RT 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 secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from ) diluted to 1:10 000 in TBS-T for 1h at RT with agitation. The blot was washed as above, the last wash in distilled water, and developed for 2 min with chemiluminescent detection reagent. Exposure time was 60 seconds.
Courtesy of Lotta von Sydow, KBC, Umeå, Sweden
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