Immunofluorescence analysis of Ubiquitin B. Staining was performed using 70% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Ubiquitin (2A1) Mouse Monoclonal Antibody at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal Secondary Antibody, Alexa Fluor 488 conjugate (Cat. No. A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade Gold Antifade Mountant with DAPI (Cat. No. S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Cat. No. R415, 1:300). Panel d represents the merged image showing cytoplasmic and nuclear localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
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. Invitrogen ubiquitin antibodies are validated for use in various applications. To see all the ubiquitin antibody offerings please click the link below.
Applications
Quality Invitrogen ubiquitin antibodies are available for a variety of research needs.
Immunohistochemical analysis of ubiquitin in paraffin-embedded human lung carcinoma using an ubiquitin polyclonal antibody (Cat. No. PA5-17067).
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For Research Use Only. Not for use in diagnostic procedures.
For Research Use Only. Not for use in diagnostic procedures.