ANN-1 | Annexin-1
AS18 4166 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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Recombinant Ann1 (1), Ann4 OE (2). Leaves of Arabidopsis thaliana were ground in liquid nitrogen and total protein was extracted using extraction buffer: Tris pH-8.0 (1M) 5%, EDTA pH-8.0 (0.5 M) 0.2%, β-Mercaptoethanol 0.05%, PMSF (1%) 0.05%, Triton X-100 1%, in distilled water. For each sample, 50 µg of extracted protein was taken, diluted with distilled water and added with 4xLaemli Sample buffer. The samples were denatured at 95℃ for 5 min and were separated on 12 % SDS-PAGE. The samples were blotted for 2 h to PVDF membrane using tank/wet transfer at 4℃ with constant stirring. The blots were blocked with blocking solution (5% dried milk in 1xTBS) for overnight at 4℃ with agitation. Blots were incubated in the primary antibody at a dilution of 1:1000 for 1h at RT with agitation in 5% dried milk in 1xTBS. The antibody solution was decanted and the blots were rinsed briefly, then washed once for 15 min and 3 times for 5 min in 1xTBS at RT with agitation. Blots were incubated in secondary antibody (Goat anti-rabbit IgG (HRP conjugated)) diluted to 1:50 000 in 5% dried milk in 1xTBS for 1h at RT with agitation. The blots were washed as mentioned above and developed for 5 min with chemiluminescent detection reagent in dark room. The exposure time was 10 min.
Courtesy Dr. dr Umesh Tanwar, Departament of Plant Biochemistry Institute of Biochemistry and Biophysics PAS, Poland
Alternative names: ANNAT1, ANX23-ATH, ATOXY5, OXY5, Annexin D1, AnnAt1.
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