DCL3 | Dicer-like protein 3
AS12 2103 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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50 µg of total protein from Arabidopsis thaliana total cell extract which came from the ground, frozen powder and has been directly transferred to 2x Laemmli sample buffer was separated on 15 % SDS-PAGE and blotted over night at 64 mA to nitrocellulose (wet transfer). Blots were blocked with Roti-block over night at 4°C agitation. Blot was incubated in the primary antibody at a dilution of 1: 2000 (1) and 1: 500 (2) 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 times for 5 min in PBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602) diluted to 1:10 000 for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturers instructions. Exposure time was 1 min.
Courtesy of Dr. Sascha Laubinger, ZMBP, Germany
Recommend protein load is from 40-50 µg/well (using 1.5 mm spacers helps to obtain wider wells)
DCL3 (EC=3.1.26) is a ribonuclease (RNase) III involved in RNA-mediated post-transcriptional gene silencing (PTGS). Functions in the microRNAs (miRNAs) biogenesis pathway by cleaving primary miRNAs (pri-miRNAs) and precursor miRNAs (pre-miRNAs).
Synonymes: Endoribonuclease Dicer homolog 3.