Rabbit Ubiquitin Polyclonal Antibody

Anti-Ubiquitin Antibody

Cat. Num. AAA4158617

• Applications: Western Blot, Immunohistochemistry, ELISA
• Synonyms: Ubiquitin; Polyclonal Antibody; Anti-Ubiquitin Antibody; anti-Ubiquitin antibody

• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Specificity: This product is an IgG fraction antibody purified from monospecific antiserum by a multi-step process which includes delipidation, salt fractionation and ion exchange chromatography followed by extensive dialysis against the buffer stated above. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Rabbit Serum.
• Form/Format: Liquid; PBS, pH 7.4 with 0.05% sodium azide.
• Concentration: 1ug/ul (varies by lot)
• Sequence Length: 128

• Applicable Applications for anti-Ubiquitin antibody: Western Blot (WB), Immunohistochemistry (IHC), ELISA (EIA)
• Application Notes: This purified polyclonal antibody reacts with ubiquitin by ELISA and western blot. Although not tested, this antibody is likely functional in immunohistochemistry and immunoprecipitation. For detection of free Ub by western blotting use Tris-Tricine SDS-PAGE and autoclaved nitrocellulose filters after the transfer and before blocking and addition of anti-Ub antibodies. Details on western blotting procedures are found in Mimnaugh et al., (1999 and 2002).
• Immunogen: This purified antibody was prepared from rabbit serum after repeated immunizations with ubiquitin coupled to rabbit IgG using glutaraldehyde.
• Preparation and Storage: This product is stable for several weeks at 4 degree C as an undiluted liquid. Dilute only prior to immediate use.; For extended storage, aliquot contents and freeze at-20 degree C or below. Avoid cycles of freezing and thawing. Expiration date is one (1) year from date of receipt.

Testing Data

Related Product Information for anti-Ubiquitin antibody

Ubiquitin (Ub) is a small, 76-residue, protein (8.5 kDa) found both as free monomer and covalently attached to itself and other proteins in eukaryotic cells. Free Ub is a very compact and stable molecule that is easily refolded after being denatured. It is therefore recommended that for detection of free Ub on Westerns, the Tris-Tricine SDS-PAGE is used and nitrocellulose filters are autoclaved after the transfer and before blocking and addition of anti-Ub antibodies. The Cterminus of ubiquitin forms an isopeptide bond with the eta-amino group of a lysine side chain in a target protein. In this way proteins can be covalently modified by the addition of ubiquitin which may alter the target protein's function. Monoubiquitination generally targets proteins for internalization, endocytosis and lysosomal degradation, or modifies the surface charge of histones and affects chromatin compaction. Conjugation of ubiquitin (Ub) involves a three-step mechanism whereby specific enzymes (or enzyme complexes) activate and covalently link Ub to their substrates. Multi-ubiquitin chains can be built up on a single lysine of the target protein, by isopeptide bond formation usually between the carboxyl group of Gly76 of one Ub with the amino group of the side chain of Lys of the preceding ubiquitin. Although Lys48 of Ub is most frequently used for multi-ubiquitin chain formation, other lysines have also been shown to form bonds with Gly76. If a chain of multiple copies of ubiquitin is attached to a protein, this appears to target the protein for degradation by the large intacellular protease known as the 26S proteasome. The process of intracellular proteasomal proteolysis is very rapid and efficient. This makes detection of certain ubiquitinated proteins extremely difficult. Several specific inhibitors of proteasomes are commercially available and have been proven to be very useful in research applications. The detection of ubiquitinated species is further complicated by the fact that the process of ubiquitination is reversible and is accomplished by highly specific proteases present in all eukaryotic cells. It is therefore recommended that cell lysates are kept on ice and the lysis buffers contain one or to be very useful in research applications. The detection of ubiquitinated species is further complicated by the fact that the process of ubiquitination is reversible and is accomplished by highly specific proteases present in all eukaryotic cells. It is therefore recommended that cell lysates are kept on ice and the lysis buffers contain one or more of cysteine protease inhibitors (n-ethylmaleimide, E64, etc.).

NCBI and Uniprot Product Information

• NCBI GI #: 295039418
• NCBI Official Full Name: ubiquitin
• Protein Family: Ubiquitin