Sec6 | Exocyst complex subunit
AS13 2686 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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50 µg of total protein from Arabidopsis thaliana, wt whole plant extract of one week old seedlings (1), whole plant extract of one week old seedlings of Arabidopsis thaliana Col-0 expressing SEC6-GFP (2), whole plant extract of one week old seedlings of Arabidopsis thaliana sec8 k.o. mutant complemented by SEC8-GFP (3), extracted with SEC6/8 buffer (20mM HEPES, pH 6.8; 150mM NaCl; 1mM EDTA; 1mM DTT; 0.5% Tween 20) were separated on 10 % SDS-PAGE then using semi-dry transfer blotted 1h to the nitrocellulose membrane (1.5 mA/cm2). Blots were blocked with 5 % non fat milk in PBS overnight at 4°C with agitation. Blot was incubated in the primary antibody in 5 % milk in PBS at a dilution of 1: 7500 for 1h at RT with agitation. The antibody solution was decanted and the blot was rinsed five times 5 min with PBST (PBS + 0.5% Tween 20) at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:15 000 in 5 % milk in PBS for 1h at RT with agitation. The blot was washed as above and developed for 5 min with chemiluminescent detection reagent according to the manufacturer's instructions. Exposure time was 10 seconds.
Courtesy of Dr. Michal Hála, UEB, Czech Republic
Sec6 is a member of the exocyst complex involved in tethering vesicles to the plasma membrane during regulated or polarized secretion.
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