AGO5 | Argonaute 5

AS10 671 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

Product Information

Immunogen

peptide of Arabidopsis thaliana AGO5 UniProt: Q9SJK3, TAIR:AT2G27880

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 Immunofluorescebce (IF), Immunoprecipitation (IP), Western blot (WB)

Recommended dilution 1 : 100 (IF), 5 ug per gram floral tissue (IP), 1 : 1500 (WB)

Expected | apparent MW

111 | 111 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana

Predicted reactivity

Arabidopsis thaliana

Not reactive in

Zea mays

Application examples

Application examples Application example

western blot using anti-AGO5 antibodies on Arabidopsis thaliana

Arabidopsis thaliana total protein extracted by TCA-acetone precipitation (check protocol tab for details) from floral buds of Col-0 (which should be more enriched in AGO5) was saturated in 8M urea were separated on 10% SDS-PAGE and blotted for 1hour to 0.2 µm nitrocellulose at 100V using wet transfer system. Blots were blocked with 0.5% cold fish gelatin for 1hr at room temp with agitation. Blot was incubated in the primary antibody a dilution of 1:250 overnight at 4C 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 30min at RT with agitation and washed 1X with TBSTT for 15min, 1X with TBST for 15min before scanning with the ODyssey IRD scanner.

Courtesy of Dr. Betty Chung, Dr Pawel Baster, University of Cambridge, United Kingdom

Additional information

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.

Related products

collection of antibodies to micro RNA

Plant protein extraction buffer

Background

AGO5 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). Probably involved in antiviral RNA silencing.