Ubiquitin is a highly conserved protein sized around 8.5 kDa. It has an ATP-dependent role in the targeting of proteins for proteolytic degradation by the 26S proteosome. Proteolytic degradation is critical to the maintenance of appropriate levels of short-lived and regulatory proteins. These proteins include those involved in cellular metabolism, heat shock and stress response, antigen presentation, modulation of cell surface receptors and ion channels, cell cycle regulation, transcription, and signaling factors.
The ubiquitin-proteasome pathway deconstructs most proteins in the eukaryotic cell cytoplasm and nucleus. To perform this function, the protein to be degraded is first covalently attached to the C terminus of ubiquitin, and the ubiquitinated complex is then recognized by a complex of degradative enzymes.
Interestingly, ubiquitin also becomes covalently bonded to many types of pathological inclusions, which appear to be resistant to normal degradation. Therefore, ubiquitin antibodies are very useful for studies of these inclusions. For example, the neurofibrillary tangles and paired helical filaments diagnostic of Alzheimer's disease, Lewy bodies seen in Parkinson's disease, and Pick bodies found in Pick's disease are all heavily ubiquitinated and can be readily visualized with ubiquitin antibodies. Quality Invitrogen ubiquitin antibodies are available for a variety of research needs.
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Immunofluorescent analysis of ubiquitin (green) in HeLa cells either left untreated (left panel) or treated with 50µM of MG132 proteasome inhibitor (right panel) for 90 minutes. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Thermo Scientific Blocker BSA (Cat. No. 37525) for 15 minutes at room temperature. Cells were probed with a ubiquitin monoclonal antibody (Cat. No. MA1-10035) at a dilution of 1:200 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488-conjugated goat-anti-mouse IgG secondary antibody (Cat. No. 35502) at a dilution of 1:400 for 30 minutes at room temperature. F-actin (red) was stained with DyLight 554-conjugated phalloidin (Cat. No. 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Cat. No. 62249). Images were taken on a Thermo Scientific ArrayScan imager at 20x magnification.
For Research Use Only. Not for use in diagnostic procedures.