AGO10 | Argonaute 10
AS15 3071 | clonality: polyclonal | host: rabbit | reactivity: Arabidopsis thaliana
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|Recommended dilution||1 : 10 000 (WB)|
|Expected | apparent MW||
|Confirmed reactivity||Arabidopsis thaliana|
A. lyrata, B. napus, C. rubella, C. clementina, C. sinensis, E. salsugineum, G. arboreum, G. raimondii
|Not reactive in||
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.
|Selected references||To be added when available, antibody released in April 2016.|
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 Amersham ECL Prime and expose to Amersham Hyperfilms ECL for 5 minutes.
Courtesy of Dr. Dr. Pablo Manavella, Instituto de Agrobiotecnología del Litoral (IAL), Argentina
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