Anti-Linear Ubiquitin, clone 1E3 Antibody

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.

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.

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

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.

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.