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

product information

AGO1 belongs to a group of argonaute proteins which are catalytic component of the RNA-incudes silencing complex (RISC). This protein complex is responsible for the gene silencing (RNAi).

N-terminal peptide of Arabidopsis thaliana AGO1 O04379

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), immuprecipitation (IP)

AS09 617  | AGO4 argonaute 4, rabbit antibody

AS10 672 | AGO6 | argonaute 6, rabbit antibody

AS10 673 | AGO9 | argonaute 9, rabbit antibody

collection of antibodies to micro RNA

Recommended secondary antibody for ECL detection

antibody binds microRNA and tasiRNAs, preference for 21nt miRNAs with 5'U

application information

1: 5000 - 1: 10 000 (ECL Plus), 5 µg of antibody per gram of tissue (IP)

116.4 | 130  kDa

Arabidopsis thaliana

Pisum sativum, Ricinus communis, Vitis vinifera

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. 

The AGO1 antibody is extremely specific to AGO1 and does not cross-react with other antibodies. The evidence is 1) the peptide to which it was raised is at the very N-terminus of the protein and is not present in other AGOs 2) aAGO1 does not cross react with the AGOs which are overexpressed (AGO2, AGO3, AGO4, AGO5, AGO6, AGO9) using a western blot.

Baumberger et al. (2007). The polerovirus silencing suppressor PO targets ARGONAUTE proteins fo degradation. Current Biology 17: 1609-1614.

application example

80 µg of Arabidopsis thaliana soluble total cell extract (extracted in 20mMTris 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-AGO1 antibody (1:10 000, 1h) andsecondary anti-rabbit (1:10000, 1 h) antibody (HRP conjugated, Santa Cruz(sc-2054)) in TBS-TT containing 5% low fat milk powder. Antibody incubationswere followed by washings in TBS-TT. All steps were performed at RT withagitation. Blots were developed for 5 min with ECL-PLUS detection reagent according the manufacturer's instructions (GE Healthcare). Exposure time was 5 seconds.

Courtesy Dr. Ericka Havecker, University of Cambridge