Ubiquitin

Ubiquitin is a highly conserved globular 76 amino acid protein of about 8.5kD. It has an important role in the targeting of proteins for proteolytic degradation. Proteins to be degraded are covalently coupled to the C-terminus of ubiquitin by means of ubiquitin ligases. The ubiquitin itself is frequently also ubiquitinated, producing a polyubiquitin chain. The polyubiquitinated complex is then recognized by a complex of degradative enzymes which form the proteosome. Ubiquitin is also covalently attached to a variety of other pathological inclusions seen in human diseases which appear to be resistant to normal degradation. These inclusions include the neurofibrillary tangles of Alzheimer's disease and progressive supranuclear palsy, Pick bodies of Pick's disease, Lewy bodies of Parkinson's disease, Mallory bodies of alcoholic liver disease, Rosenthal fibers of Alexander's disease, and the inclusion bodies in inclusion myositis and oculopharyngeal muscular dystrophy. The ubiquitin molecule appears to be present in all eukaryotic cells and has an identical primary structure in all animals. Ubiquitin is present in the nucleus, cytoplasm and on cell surface membranes.\n\nApplications:\nCan be used for immunoassays, Western Blot, Immunofluorescence and Immunoprecipitation. Furthermore the antibody is useful for Immunohistology after heat-induced retrieval on both frozen and paraffin embedded sections.\n\nRecommended Dilution:\nImmunohistology: 1:10\nImmunofluorescence: 1:10\nWestern Blot: 1:10. For Western Blot it is recommended to use lysis buffer in 10mM N-ethylmaleimide to inhibit ubiquitin-conjugating enzymes. N-ethylmaleimide inactivates certain enzymes by blocking free sulfhydryls. After eletrophoresis and transfer, pre-incubate transferred membranes in denaturing buffer (6M guanidine-HCl, 20mM Tris-HCl, 5mM beta-mercaptoethanol, 1mM PMSF (pH 7.5)) for 30-60 minutes at 4 degrees C, followed by extensive PBS washing.\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%). Store at -20 degrees C or colder. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

U1000-04C

Size

100ug

Applications

IF, IHC, IP, WB

Hosts

Mouse

Reactivities

Hum, Bov, Ch/Bird, Dro/Arth

Form

Supplied as a liquid in PBS, containing 0.02% sodium azide and 0.1% BSA.

P Type

Mab

Purity

Purified by Protein G affinity chromatography.

Isotype

IgG1

References

1. Sternsdorf, T et al., (1999) Mol Cell Biol 19:5170. 2. Holbert, S et al., (2001) Proc Natl Acad Sci USA 98:1811. 3. Mullally, J et al., (2001) J Biol Chem 276:30,366.

Additional Info

Recognizes ubiquitin, both conjugated and unconjugated. Reactivity has been shown in human, bovine, chicken, Drosophilia and C. elegans.