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AGO10 | Argonaute 10

AS15 3071 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

AGO10 | Argonaute 10 in the group Antibodies for Plant/Algal  / DNA/RNA/Cell Cycle / microRNA at Agrisera AB (Antibodies for research) (AS15 3071)

DATA SHEET IN PDF

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Product name, number (Agrisera, Sweden)

Data sheet Product citations Protocols Add review

Product Information

Immunogen

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana AGO10 protein sequence, Uniprot: Q9XGW1, TAIR: AT5G43810

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 Western blot (WB)
Recommended dilution 1 : 10 000 (WB)
Expected | apparent MW

110.9 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana
Predicted reactivity

A. lyrata, B. napus, C. rubella, C. clementina, C. sinensis, E. salsugineum, G. arboreum, G. raimondii. N. benthamiana


Species of your interest not listed? Contact us
Not reactive in

Zea mays

Application examples

Application examples Application example

western blot using anti-AGO10 antibodies

50 µg of total protein from Arabidopsis thaliana inflorescences were extracted with extraction buffer (50 mM Tris pH7.5; 150 mM NaCl; 1 mM EDTA; 10 % v/v Glycerin; 1 mM DTT, 1x Complete Protease Inhibitor Cocktail, Roche) and denatured with Laemmli buffer at 95ºC 5 min. were separated on 10% SDS-PAGE and blotted 1.5 h to PVDF using tank transfer. Blots were blocked with blocking buffer (3% milk powder; 1x TBS; 0.1% Tween-20) 1 h at RT with agitation. Blot was incubated in the primary antibody at a dilution of 1:10 000 ON at 4ºC with agitation. The antibody solution was decanted and the blot was rinsed briefly and then washed tree times for 15 min. in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602) diluted to 1:20 000 in blocking buffer for 1h at RT with agitation. The blot was washed as above and developed for 5 min with chemiluminescent detection reagent of extreme femtogram sensitivity, exposed to Amersham Hyperfilms ECL for 5 minutes.

Courtesy of Dr. Dr. Pablo Manavella, Instituto de Agrobiotecnología del Litoral (IAL), Argentina

Additional information

AGO expression may be cell/tissue specific and using floral tissue is recommended where most of the AGOs are expressed the highest. Seedlings can be used as a negative control.

Use of proteasome inhibitors as MG132 can help to stabilize AGO proteins during extraction procedure.

Related products

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collection of antibodies to micro RNA

Plant protein extraction buffer

Secondary antibodies

Background

Background

AGO10 is involved in miRNA binding, and in RNA-mediated posttranscriptional gene silencing (PTGS).

Product citations

Selected references Sprunck et al. (2019). Elucidating small RNA pathways in Arabidopsis thaliana egg cells.

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