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product information
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| background |
|
AGO6 (Argonaute 6) 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). |
| immunogen |
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KLH-conjugated peptide derived from Arabidopsis thaliana AGO6 At2g32940 |
| antibody format |
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rabbit |
polyclonal |
|
affinity purified serum |
lyophilized |
|
| quantity |
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100 µg |
for reconstitution add 100 µg of sterile water. |
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| storage |
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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 |
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western blot (WB) |
| related products |
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AS09 527 | AGO1 | argonaute 1 AS09 617 | AGO4 argonaute 4, rabbit antibody AS10 673 | AGO9 | argonaute 9, rabbit antibody collection of antibodies to micro RNA Recommended secondary antibody for ECL detection |
| additional information |
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There can be a reaction with NEB broad range prestained protein ladder depending upon secondary antibody used. AGO6 accumulates to lower levels in the nrpd1a mutant (Havecker at el. 2010). In addition, a non-specific band migrates to a similar location and thus protein was run on a 6% denaturing gel to obtain good separation. If other secondary antibody is used than in the figure below, cross-reacting pattern can be different. |
application information
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| recommended dilution |
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1: 1000 (WB) |
| expected | apparent MW |
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116.4 | 99 |
| confirmed reactivity |
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Arabidopsis thaliana |
| predicted reactivity |
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Arabidopsis thaliana |
| not reactive in |
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no confirmed exceptions from predicted reactivity known in the moment |
| additional information |
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antibody binds microRNA and tasiRNAs, preference for 24nt siRNAs with 5'U 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. |
| selected references |
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Havecker, E.R., Wallbridge, L.M., Hardcastle, T.J., Bush, M.S., Kelly, K.A., Dunn, R.M., Schwach, F., and Baulcombe, D.C. The RNA-directed DNA methylation Arabidopsis Argonautes functionally diverge based on expression and interaction with target loci. manuscript revisions submitted, awaiting acceptance. |
80 µg of Arabidopsis thaliana soluble total cell extract (1), Arabidopsis thaliana nrpd1a mutant which accumulates lower levels of AGO6 (2), were extracted in 20m M Tris pH7.5, 5mM MgCl2, 2.5mM DTT, 300mM NaCl, 0.1% NP-40, 1% protease inhibitor. Proteins were 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-AGO6 antibody, AS10 672 (1:1000, 1h) and secondary anti-rabbit (1:15000, 1 h) antibody (HRP conjugated, Agrisera AS09 602) 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 ECL-PLUS detection reagent according the manufacturer's instructions (GE Healthcare) using Hyperfilm ECL (GE Healthcare).Exposure time was 1 min. Courtesy Dr. Ericka Havecker, University of Cambridge
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