ATG7 | Ubiquitin-like modifier-activating enzyme atg7
AS15 3061 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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1: 500 - 1: 5000 (WB)
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50 µg of the soluble proteins (Sol) were extracted from Arabidopsis thaliana leaves with extract buffer without detergent (50mM Tris-HCl pH7.5, 150mM NaCl, 1mM EDTA). The proteins were denatured with sample buffer and boiling at 95°C for 5 min and 50 ug of proteins were separated on 10% SDS-PAGE and blotted for 1h to PVDF membrane using semi-dry transfer. The blot was blocked with 0.5 % milk for 1h/RT and washed in TBS-T for 5 minute twice. The blot was incubated in the primary antibody at a dilution of 1:5 000 in Can Get Signal solution for ON/4°C. The antibody solution was decanted and the blot was rinsed briefly three times, then washed once for 15 min in TBS-T at RT with agitation. The blot was incubated in Agrisera matching secondary antibody (anti-rabit IgG horse radish peroxidase conjugated AS09 602) diluted to 1:25 000 in TBS-T for 1h/RT with agitation. The blot was washed as above and developed for 3 min with Agrisera ECL SuperBright.
Exposure Courtesy Dr Shino Goto-Yamada, Malopolska Centre of Biotechnology (MCB) Jagiellonian University, Krakow, Poland
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