AGO2 | Argonaute 2

AS13 2682 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana

Product Information

Immunogen

KLH-conjugated synthetic peptide, derived from Arabidopsis thaliana AGO2 protein, UniProt: Q9SHF3, TAIR: AT1G31280

Host Rabbit

Clonality Polyclonal

Purity Affinity purified serum in PBS, pH 7.4

Format Lyophilized

Quantity 50 µg

Reconstitution For reconstitution add 50 µl of sterile water.

Storage 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.

Tested applications Immunoprecipitation (IP), Western blot (WB)

Recommended dilution 1 : 250-1 : 500 (WB)

Expected | apparent MW

113 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana

Predicted reactivity

Capsella rubella, Solanum tuberosum

Species of your interest not listed? Contact us

Not reactive in

Chlamydomonas reinhardtii, Nicotiana benthamiana, Solanum lycopersicum, Zea mays

Application examples

Application examples Application example

western blot using anti-AGO2 antibodies

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 (check protocol tab) 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 (buffered in TBS) 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 (goat anti-rabbit DyLight® 800 conjugated, AS12 2460, Agrisera) 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

Additional information

AGO2 protein is strongly induced by stress.

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.

Antibody incubation should be done over night in 4°C. Use of material with enriched AGO2 levels is recommened.

Related products

collection of antibodies to micro RNA

Plant protein extraction buffer

Background

AGO2 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). AGO2 is probably involved in antiviral RNA silencing.

Product citations

Selected references Wang et al. (2019). The PROTEIN PHOSPHATASE4 Complex Promotes Transcription and Processing of Primary microRNAs in Arabidopsis. Plant Cell. 2019 Feb;31(2):486-501. doi: 10.1105/tpc.18.00556.
Dalmadi et al. (2019). AGO-unbound cytosolic pool of mature miRNAs in plant cells reveals a novel regulatory step at AGO1 loading. Nucleic Acids Res. 2019 Aug 8. pii: gkz690. doi: 10.1093/nar/gkz690.
You et al. (2019). FIERY1 promotes microRNA accumulation by suppressing rRNA-derived small interfering RNAs in Arabidopsis. Nat Commun. 2019 Sep 27;10(1):4424. doi: 10.1038/s41467-019-12379-z. (immunoprecipiation)