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

product information

AGO4 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).

KLH-conjugated synthetic peptide chosen from Arabidopsis thaliana AGO4 sequence Q9ZVD5 At2g27040

store at -20°C; Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur adhering to the cap or sides of the tubes.

western blot (WB), immunoprecipitation (IP), immunocytochemistry (ICC)

AS09 527 | AGO1 | argonaute 1

collection of antibodies to micro RNA

Antibody concentration is 0.4 mg/ml in 50% glycerol, PBS, 0.005% thimerosal, 0.05% BSA, threfore the antibody sample will not freeze in -20°C.

binds endogenous siRNAs, preference for 24nt siRNAs with 5' A.

application information

1: 4000 (WB), 5 µg of antibody per 1 gram of a fresh tissue (IP), 1: 100 (ICC)

103 kDa

Arabidopsis thaliana, Hyacinthus orientalis

Arabidopsis thaliana

Solanum lycopersicum, Brassica oleracea

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. 

Note that the AGO4 antibody reacts with the NEB prestained protein marker.

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.

Havecker, E.R., Wallbridge, L.M., Hardcastle, T.J., Bush, M.S., Kelly, K.A., Dunn, R.M., Schwach, F., and Baulcombe, D.C. The RNA-directed DNA methylation Arabidopsis Argonautes functionally diverge based on expression and interaction with target loci. submitted

application example

80 µg of Arabidopsis thaliana soluble total cell extract (2) (extracted in 20m MTris 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-AGO4 antibody (1:4000, 1h) and secondary anti-rabbit (1:15000, 1 h) antibody (HRP conjugated, Agrisera AS09 602) 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.

Note that the antibody reacts with NEB broad range prestained protein ladder (1)

Courtesy Dr. Ericka Havecker, University of Cambridge