NodGS | Nodulin / glutamate-ammonia ligase-like protein
AS15 3030 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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94 | 90 kDa
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4,5 of total protein (supernatant after 1h at 25.000 xg) from Arabidopsis thaliana suspension culture extracted with 50mM HEPES pH 7,5/75mM NaCl/1mM EGTA/1mM MgCl2/1mM NaF in the presence of protease inhibitors were separated on 8 % SDS-PAGE and blotted 1,5h to a nitrocellulose membrane using tank transfer. Blots were blocked with 5% milk for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 500, 1:1000, 1:2.000, and 1:5000 overnight at 4°C without agitation. The antibody solution was decanted and the blot was rinsed, then 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 1:10 000 in 5% milk for 30 min at RT with agitation. The blot was washed as above and developed for 1 min with chemiluminescent detection reagent. Exposure time was 15 seconds.
Courtesy of Dr. Jana Chumová, Laboratoř funkční cytologie, Czech Republic
NodGS (Nodulin/glutamate-ammonia ligase-like protein) shares homology with nodulins and self-assembles into oligomers which are present in response to flagellin treatment. It contains a C-terminal domain of a prokaryotic glutamine synthetase type I. Protein is mainly localized to cytoplasm and is present in a oligomeric form of ca. 700 kDa.