UBQ11 | Ubiquitin (affinity purified)
AS08 307A | Clonality: Polyclonal | Host: Rabbit | Reactivity: A.thaliana, H.vulgare, L. angustifolius, M.domestica, N. benthamina, O. sativa, P. sativum, S.tuberosum, Z.mays
|Recommended dilution||1 : 10 000 (WB)|
|Expected | apparent MW||
|Confirmed reactivity||Arabidopsis thaliana, Hordeum vulgare, Lupinus angustifolius, Malus domestica, Nicotiana benthamina, Oryza sativa, Pisum sativum, Solanum tuberosum, Zea mays
|Predicted reactivity||Brachypodium distachyon, Brassica napus, Capsella rubella, Cochliobolus heterostrophus, Citrus clementina, Citrus sinensis, Coffea canephora, Cymbidium faberi, Eucalyptus grandis, Erythrane guttata, Glycine max, Glycine soja, Ipomoea batatas, Medicago truncatula, Magnaporthe oryzae, Nicotiana tabacum, Phaseolus vulgaris, Populus trichocarpa, Prunus persica, Pyrus communis, Ricinus communis, Solanum nigrum, Sorghum bicolor, Triticum aestivum, Theobroma cacao, Vitis vinifera|
|Not reactive in||Algae|
Antibody can be used to check qubiquitination status in the whole plant extracts.
|Selected references||Krasuska et al. (2016). Nitric oxide-polyamines cross-talk during dormancy release and germination of apple embryos. Nitric Oxide. 2016 Nov 23. pii: S1089-8603(16)30178-1. doi: 10.1016/j.niox.2016.11.003. [Epub ahead of print]|
The 10 μg of soluble protein extracts from 3 -week old Solanum tuberosum (1), one -week old seedlings of Lupinus angustifolius (2), Pisum sativum (3) , Zea mays L . (4) and Hordeum vulgare (5) leaves were extracted with buffer containing 50 м M Hepes- KOH, pH 7.5, 330 м M sorbitol, 2 м M EDTA, 1 м M MgCl 2, 5 м M ascorbate, 0.05% BSA . All samples were mixed with sample buffer, denatured for 5 min at 70°C and separated on 10% Tricine -SDS -PAGE . Proteins were blotted 30 min to HybondTM -PVDF membrane (Amersham Life Science) using semi -transfer (Bio -Rad) in standard transfer buffer in presence of 20% methanol. Transfer of proteins to the membrane was checked using 1% Ponceau S staining before the blocking step. Blots were blocked in buffer (5% low -fat milk in 1xPBS, 0. 1% Tween) with for 60 min at room temperature (RT) with agitation. Blots were incubated in the primary antibody at a dilution of 1: 5 000 at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in P BS -T at RT with agitation. Blot was incubated in secondary antibody
(goat anti -rabbit IgG, AS09 602, Agrisera ) diluted to 1 :50 000 for 1h at RT with agitation. The blot was washed as above and developed for 5 min with Clarity Western ECL Substrate and ChemiDoc detection system (Bio-Rad).
Total cell extracts of Solanum tuberosum (1), Lupinus angustifolius (2), Pisum sativum (3), Zea mays L. (4), Hordeum vulgare (5), were separated in conditions as described above. Marker - P restained SDS -PAGE ( Bio Rad, kat # 1610318). Monomers are marked by arrow. Exposure 30 seconds. For visualization of monomers, longer exposure time was applied.
Courtesy of Dr. Elena Poznidaeva Laboratory of Plant Gene Expression, Timiryazev Institute of Plant Physiology RAS, Moscow, Russia
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