UBQ11 | Ubiquitin (affinity purified)

Ubiquitin mutant is lethal, therefore could not be used in validation of this antibody.,Antibody can be used to check qubiquitination status in the whole plant extracts.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.Suggested extraction buffer: 100 mM Tris-HCl, pH 8.0, 0.1% [w/v] SDS, 0.5% [w/v] sodium deoxycholate, 1% [v/v] glycerol, 50 mM sodium metabisulfite, 20 mM N-ethylmaleimide (NEM) and protease inhibitor cocktail (Roche) (Orosa et al. 2018). This buffer will help to stabilize the conjugates and will help to detect any increase or decrease in conjugate accumulation using the antibodies.

Ubiquitin is a highly conserved regulatory protein that is expressed in all eukaryotic tissues. Initially, the protein is called ubiquitous immunogenerative polypeptide. Its function is to label proteins for degradation by the ubiquitin proteasome system (UPS).