AGO9 | Argonaute 9
AS10 673 | clonality: polyclonal | host: rabbit | reactivity: Arabidopsis thaliana
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1: 10 000 (WB), 5 µg of antibody per 1 gram of a fresh tissue (IP)
|Expected | apparent MW||
|Not reactive in||
no confirmed exceptions from predicted reactivity known in the moment
AGO expression may be cell/tissue specific and using floral tissue is recommended where most of the AGOs are expressed the highest. Seedlings can be used as a negative control.
|Selected references||Havecker et al. (2010) The RNA-directed DNA methylation Arabidopsis Argonautes functionally diverge based on expression and interaction with target loci. Plant Cell 22(2): 321-34.|
80 µg of Arabidopsis thaliana soluble total cell extract (extracted in 20 mMTris pH7.5, 5mM MgCl2, 2.5mM DTT, 300mM NaCl, 0.1% NP-40, 1% proteaseinhibitor) was separated on 6% SDS-PAGE and blotted 1h to nitrocellulose. Filters were blocked 1h with 5% low-fat milk powder in TBS-TT (0.25% TWEEN20; 0.1% Triton-X) and probed with anti-AGO9 antibody (1:10 000, 1h) and secondary anti-rabbit antibody (HRP conjugated, Agrisera AS09 602) (1:15 000, 1 h) in TBS-TT containing 5% low fat milk powder. Antibody incubations were followed by washings in TBS-TT. All steps were performed at RT with agitation. Blots were developed for 5 min with ECL-PLUS detection reagent according the manufacturer's instructions (GE Healthcare). Exposure time was 30 seconds.
Courtesy Dr. Ericka Havecker, University of Cambridge
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