ERD7 | Early Response to Dehydration 7
AS19 4317 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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Ten µg of total protein extracted freshly from Arabidopsis thaliana total leaf with protein extraction buffer (Tris-HCl 50 mM pH 8.0, 150 mM NaCl, 1% Triton X-100, 0.1% SDS, 0.5% Na-Deoxycholate, 2 mM PMSF, 2 mM DTT) and denatured with Laemmli sample buffer containing 2% beta-mercaptoethanol at 70°C for 10 min. Samples were separated on 10% SDS-PAGE and blotted 1h to PVDF using semi-dry transfer. Blot was blocked with 5% non-fat milk for 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 2 000 in TBS-T with 1% milk for ON/4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly, then washed 3 times for 5-10 min in TBS-T at RT with agitation. Blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated AS09 602) diluted to 1:25 000 in TBS-T with 1% milk for 1h/RT with agitation. The blot was washed as above and developed for 5 min with chemiluminescent detection reagent according to manufacture's instructions. Exposure time was 1 min.
Courtesty Dr. Hiroaki Fujii, University of Turku, Finland
The protein is detected in the microsomal fraction. (Barajas-Lopez et al, 2021)
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