UBQ11 | Ubiquitin
AS08 307 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, H. vulgare, N. benthamina, P.sativum, S. lycopersicum
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5 μg of recombinant protein from Phytophora sp. (1), human (2), Arabidopsis thaliana His-tagged ubiquitin (3), Arabidopsis thaliana His-tagged SUMO protein (4), was separated on 15% PAA gel and blotted on PVDF membrane. Filters were blocked in 5% milk for 1h, incubated with 1: 10 000 anti-ubiquitin antibody (1h), followed by incubation with 1: 15 000 secondary anti-rabbit antibodies(1h) coupled with HRP and visualization (10 seconds exposure) with standard chemiluminescent detection reagent.
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.
Ubiquitin is a highly conserved regulatory protein expressed in all eukaryotic tissues. Originally this protein was called: Ubiquitous Immunopoietic Polypeptide. Its function is labeling of proteins for degradation through ubiquitin proteasome system (UPS).
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