SAM1-4 | S-adenosylmethionine synthase
AS16 3148A | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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|Recommended dilution||1: 2000 (WB)|
|Expected | apparent MW||
43.2 | 45 kDa (Arabidopsis thaliana)
|Confirmed reactivity||Arabidopsis thaliana, Malus domestica
Beta vulgaris, Brassica sp., Citrus sp., Coffea canephora, Caomelina sativa, Capsella rubella, Cucumis melo, Cucumis sativus, Genlisea aurea, Gentiana triflora, Guzmania wittmacki, Gossypium raimondii, Eucalyptus grandis, Eutrema salsugineum, Ipomoea batatas, Jatropha curcas, Musa acuminata, Nicotiana sp., Oryza brachyantha, Phoenix dactylifera, Populus sp., Prunus sp., Ricinus communis, Sesamum indicum, Setaria italica, Solanum pennellii, Solanum tuberosum, Spinacia oleracea, Tarenaya hassleriana, Theobroma cacao, Zea mays
|Not reactive in||
No confirmed exceptions from predicted reactivity known in the moment.
To be added when available, antibody released in July 2017.
70 μg of total protein from Arabidopsis thaliana wt Col-0, SAM3 knockdown, SAM1-4 knockdown, and one oeSAM3-YFP lines extracted with mortar and pestle using 2xSDS loading buffer (100 mM Tris-HCl pH 6.8, 4% SDS, 0.02% bromophenol blue, 200 mM DTT), and denatured in the same buffer at 95°C for 10 min. Samples were separated on 11% SDS-PAGE and blotted for 1h to PVDF using tank transfer. Blots were blocked with 5% milk powder in TBS-T overnight in a cold room with agitation. Blot was incubated in the primary antibody at a dilution of 1: 2 000 for 1h at RT with agitation. The antibody solution was decanted and the blot was rinsed five times for 4 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG HRP-conjugated, from Agrisera) diluted to 1: 20 000 in for 1h at RT with agitation. The blot was washed as above and developed for 2 min with Millipore Immobilon Western detection reagents. The proteins were detected using CCD Image Fusion Fx7 after 30 seconds exposure time.
Courtesy of Louis-Valentin Meteignier, LGBP, Faculté des Sciences de Luminy, France
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