UBQ11 | Ubiquitin (affinity purified)
AS08 307A | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, M. domestica, N. benthamina, O. sativa
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1: 10 000 with standard ECL (WB)
|Expected | apparent MW||
|Confirmed reactivity||Arabidopsis thaliana, Malus domestica, Nicotiana benthamina, Oryza sativa|
dicots including: Nicotiana tabacum, Glycine max, Pisum sativum, monocots including: Hordeum vulgare, Oryza sativa, Zea mays
|Not reactive in||
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]|
10 μl of a total protein from Arabidopsis thaliana wt (20 μg of total protein) and Nicotiana benthamiana, was separated on 8% polyacralymide gel and blotted on PVDF membrane. Filters were blocked in 5% milk for 1h, incubated with 1: 5 000 anti-ubiquitin antibody (3h), followed by incubation with 1: 10 000 secondary anti-rabbit antibodies (1h) coupled with HRP and visualization (1 minute exposure) with standard ECL.
Technical note: It is very difficult to detect ubiquitin monomers in total cell extracts due to a great abundance of poly and multi-ubiquitinated proteins. Recommended is size separation of protein extracts before gel electrophoresis focused on good resolution of region between 6-10 kDa.
Courtesy of Dr. Ari Sadanandom, University of Durham, UK
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