Anti-Linear Ubiquitin, clone 1E3 Antibody

Multiple molecular weight bands observed

Ubiqutin (Ub) is initially produced as a 229-a.a. Polyubiquitin-B (UniProt P0CG47) precursor protein encoded by the UBB gene (Gene ID 7314) or a 685-a.a. Polyubiquitin-C precursor protein (UniProt P0CG48) encoded by the UBC gene (Gene ID 7316) in human. The C-termini of Polyubiquitin-B (Cys229) and Polyubiquitin-C (Val685) are post-translationally removed, followed by additional posttranslational cleavages to yield multiple copies of idential 76-amino acid ubiquitin (also known as high mobility group nonhistone protein HMG-20) molecules (three copies per Polyubiquitin-B and nine copiesf per Polyubiquitin-C). Ub is involved in one of the most common post-translational modifications (PTMs) of cellular proteins, where Ub is linked covalently via its carboxyl terminus (Gly76) to usually Lys residues in target proteins. A given target Lys residue can be linked to one single Ub molecule (monoubiquitylated) or to a chain of Ub molecules (polyubiquitylated). In a polyUb chain, Ub molecules can be linked through one of the seven Lys residues (K6, K11, K27, K29, K33, K48, and K63) or through the Ub N-terminus Met1 residue (which generates linear chains). An Ub chain with a single linkage type is referred to as homotypic, whereas an heterotypic chain contains mixed linkages within the same chain, which can lead to either branched/forked (as a result of one Ub linked to two or more Ubs) or non-branched Ub chains. Lys48-linked chains mark proteins for proteasomal degradation, while Lys63-linked chains regulate the activity/function of cellular proteins.

Recombinant protein corresponding to all linear ubiquitin linkages.

Immunocytochemistry Analysis: A 1:50 dilution from a representative lot detected linear ubiquitin in Hoi-1L/Hoip transfected Hela cells.

Immunoprecipitatation Analysis: 10 µg from a representative lot immunoprecipitated linear ubiquitin from 500 µg of Hoi-1L/Hoip transfected HEK293 cells.

Western Blotting Analysis: A representative lot from an independent laboratory detected K64-linked diUbiquitin (Matsumoto, M. L., et al. (2012). J Mol Biol. 418(3-4):134-144.).

Research Category
Signaling

Research Sub Category
Cell Cycle, DNA Replication & Repair

This Anti-Ubiquitin antibody, clone 1E3 is validated for use in western blotting, IP & ICC for the detection of Linear Ubiquitin.

Format: Purified

Protein A purified

Purified rabbit monoclonal IgGκ in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Stable for 1 year at 2-8°C from date of receipt.

Evaluated by Western Blotting in linear polyubiquitin chains, K48 Ubiquitin 2-7, K63 Ubiquitin 2-7, and K11-only Ubn-ubiquitinylated HM substrate protein synthesized using K11-only mutant ubiquitin.

Western Blotting Analysis: A 1:1,000 dilution of this antibody detected linear ubiquitin chains, and not K48 Ubiquitin 2-7, K63 Ubiquitin 2-7, or K11-only Ubn-ubiquitinylated HM substrate protein synthesized using K11-only mutant ubiquitin.

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