Plant/Algal cell antibodies / DNA/RNA/cell cycle / microRNA

product information

AGO9 belongs to a group of argonaute proteins which are catalytic component of the RNA-incudes silencing complex (RISC).

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana AGO9 protein sequence Q84YI4

store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

western blot (WB), immunoprecipitation (IP)

AS09 527 | AGO1 | argonaute 1, rabbit antibody

AS09 617  | AGO4 argonaute 4,  rabbit antibody

collection of antibodies to micro RNA

to be added when available

application information

1: 10 000 (WB), 5 µg of antibody per 1 gram of a fresh tissue (IP)

101 kDa

Arabidopsis thaliana

Arabidopsis thaliana

no confirmed exceptions from predicted reactivity known in the moment

AGO expression may be tissue specific and using floral tissue is recommended where most of the AGOs are expressed the highest. Use of proteasome inhibitors as MG132 can help to stabilize AGO proteins during extraction procedure. 

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.

application example

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