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AGO9 | Argonaute 9

AS10 673 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

AGO9 | Argonaute 9 in the group Antibodies for Plant/Algal  / DNA/RNA/Cell Cycle / microRNA at Agrisera AB (Antibodies for research) (AS10 673)

DATA SHEET IN PDF

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Product Information

Immunogen

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana AGO9 protein sequence UniProt: Q84YI4, TAIR:At5g21150.

Host Rabbit
Clonality Polyclonal
Purity Affinity purified serum in PBS, pH 7.4
Format Lyophilized
Quantity 100 µg
Reconstitution For reconstitution add 100 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 Immunolocalization, whole-mount (IL), Immunoprecipitation (IP), Western blot (WB)
Recommended dilution 5 µg of antibody per 1 gram of a fresh tissue (IP), 1 : 10 000 (WB)
Expected | apparent MW

101 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana
Predicted reactivity

Arabidopsis thaliana

Not reactive in No confirmed exceptions from predicted reactivity are currently known.

Application examples

Application examples

Application example

western blot using Arabidopsis AGO9 antibodies

80 µg of Arabidopsis thaliana soluble total cell extract (extracted in 20 mMTris pH7.5, 5mM MgCl2, 2.5mM DTT, 300mM NaCl, 0.1% NP-40, 1% proteaseinhibitor) was separated on 6% SDS-PAGE and blotted 1h to nitrocellulose. Filters were blocked 1h with 5% low-fat milk powder in TBS-TT (0.25% TWEEN20; 0.1% Triton-X) and probed with anti-AGO9 antibody (1:10 000, 1h) and secondary anti-rabbit antibody (HRP conjugated, Agrisera AS09 602) (1:15 000, 1 h) in TBS-TT containing 5% low fat milk powder. Antibody incubations were followed by washings in TBS-TT. All steps were performed at RT with agitation. Blots were developed for 5 min with chemiluminescent detection reagent according the manufacturer's instructions. Exposure time was 30 seconds.

Courtesy Dr. Ericka Havecker, University of Cambridge, UK

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.

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collection of antibodies to micro RNA

Plant protein extraction buffer

Background

Background

AGO9 belongs to a group of argonaute proteins which are catalytic component of the RNA-incudes silencing complex (RISC).

Product citations

Selected references Sprunck et al. (2019). Elucidating small RNA pathways in Arabidopsis thaliana egg cells. http://dx.doi.org/10.1101/525956
Su et al. (2017). The THO Complex Non-Cell-Autonomously Represses Female Germline Specification through the TAS3-ARF3 Module. Curr Biol. 2017 Jun 5;27(11):1597-1609.e2. doi: 10.1016/j.cub.2017.05.021.
Havecker et al. (2010) The RNA-directed DNA methylation Arabidopsis Argonautes functionally diverge based on expression and interaction with target loci. Plant Cell 22(2): 321-34.

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