EPSP Synthase（3-phosphoshikimate 1-carboxyvinyltransferase, chloroplastic）に対する抗体 抗EPSP Synthase抗体

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glyphosate代謝経路で機能する酵素EPSP Synthase（3-phosphoshikimate 1-carboxyvinyltransferase, chloroplastic）に対する抗体です。

0.5 μg of purified Arabidopsis thaliana EPSPS was separated on 10 % SDS-PAGE and blotted 2h to nitrocellulose Hibond ECLmembrane (GE Healthcare) using semi-dry or tank transfer. Blots were blocked with for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody in TBS-T with (0.1 % Tween 20) at a dilution of 1: 1 000 for 1h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 2 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 Agrisera AS09 602) diluted to 1: 20 000 in for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturer's instructions. Protein detection was done by Image Quant LAS4000 (GE-Health Care) digital imager system with exposure time was of 5 minutes.
Recombinant Arabidopsis thaliana EPSPS protein (aka 3-phosphoshikimate 1-carboxyvinyltransferase) used in the assay was identified by LCMS.

Courtesy of Dr. Altanbadralt Sharkhuu, King Abdullah University of Science and Technology, Thuwal, Saudi Arabia