AGO5 | Argonaute 5
AS10 671 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

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Product Information
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

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

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.
Background
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.
Product citations
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