Agrisera/AGO2 | Argonaute 2/AS13 2682/

Antibody available in 2016 to participate in testing, please contact support@agrisera.com


product information
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.
Immunogen		KLH-conjugated synthetic peptide, derived from Arabidopsis thaliana AGO2 protein, UniProt: Q9SHF3, TAIR: AT1G31280
Host		Rabbit
Clonality		Polyclonal
Clone
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		western blot (WB)
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Additional information		TCA acetone total protein precipitation method

application information
Recommended dilution		1: 250 - 1: 500 (WB)
Expected \| apparent MW		113 kDa
Confirmed reactivity		Arabidopsis thaliana
Predicted reactivity		Capsella rubella, Solanum tuberosum
Not reactive in		Chlamydomonas reinhardtii, Solanum lycopersicum, Zea mays
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.
Selected references		To be added when available, antibody released in April 2014.

Application example

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 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 (DayLight 800®, Agrisera AS11 1827) 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

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Agrisera/AGO2 | Argonaute 2/AS13 2682/