Goat anti-Chicken IgY (H&L), DyLightŪ 800 conjugated
AS11 1827 | clonality: polyclonal | host: goat | reactivity: chicken IgY (H&L)
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1:20 – 1:2 000 (IHC), 1:50 – 1:5 000 (ICC)
to be added when available
300 µg/well of Arabidopsis thaliana protein from wilde type and AGO1-36 knock out, AGO1 knockdown mutant (1-25) were extracted by TCA-acetone precipitation from floral tissue and saturated in 8M urea were separated on 15% SDS-PAGE (1 mm thick gel) and blotted for 1hour to 0.2 µm nitrocellulose at 100V using wet transfer system. Blots were blocked with 0.5% cold fish gelatin for 1hr at room temp with agitation. Blot was incubated in the primary antibody at a dilution of 1:250 for an hour at RT with agitation. The blots were washed with 3X 15min TBS-TT at RT with agitation. Blots as incubated in the secondary antibody (DayLight 800®, Agrisera AS11 1827) 1:5000 dilution for 30 min. at RT with agitation and washed 1X with TBSTT for 15min, 1X with TBST for 15min before scanning with the ODyssey IRD scanner.
AGO2 is enriched in AGO1 knockdown mutant, which agrees with already published data Harvey et al. (2011), PLOS One.
Courtesy of Dr. Betty Chung, University of Cambridge, United Kingdom
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