The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ab189672 is ideal for use in assays that utilize avidin-linked reagents for visualization or quantitation. Reaction conditions will need to be optimized for each specific application.
>95% by SDS-PAGE . Under reducing conditions and visualized by Colloidal Coomassie® Blue stain.
Linkage specific poly-ubiquitin chains are used to investigate mechanisms of chain recognition, binding and hydrolysis by the proteasome, deubiquitinating enzymes, E3 ligases or other proteins that contain ubiquitin-associated domains (UBAs) or ubiquitin-interacting motifs (UIMs). Lys48-linked ubiquitin chains are abundant in vivo and act as a universal signal for proteasomal degradation.
For optimal results, we recommend incubation in SDS-PAGE buffer + DTT at <40 °C for 20 minutes prior to gel electrophoresis.
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Preparation and Storage
Stability and Storage
Shipped at 4°C. Store at -20°C. Avoid freeze / thaw cycle. Store In the Dark.
Constituent: 100% HEPES
This product is an active protein and may elicit a biological response in vivo, handle with caution.
Reconstitute up to 5 mg/ml
Ubiquitin exists either covalently attached to another protein, or free (unanchored). When covalently bound, it is conjugated to target proteins via an isopeptide bond either as a monomer (monoubiquitin), a polymer linked via different Lys residues of the ubiquitin (polyubiquitin chains) or a linear polymer linked via the initiator Met of the ubiquitin (linear polyubiquitin chains). Polyubiquitin chains, when attached to a target protein, have different functions depending on the Lys residue of the ubiquitin that is linked: Lys-6-linked may be involved in DNA repair; Lys-11-linked is involved in ERAD (endoplasmic reticulum-associated degradation) and in cell-cycle regulation; Lys-29-linked is involved in lysosomal degradation; Lys-33-linked is involved in kinase modification; Lys-48-linked is involved in protein degradation via the proteasome; Lys-63-linked is involved in endocytosis, DNA-damage responses as well as in signaling processes leading to activation of the transcription factor NF-kappa-B. Linear polymer chains formed via attachment by the initiator Met lead to cell signaling. Ubiquitin is usually conjugated to Lys residues of target proteins, however, in rare cases, conjugation to Cys or Ser residues has been observed. When polyubiquitin is free (unanchored-polyubiquitin), it also has distinct roles, such as in activation of protein kinases, and in signaling.
Belongs to the ubiquitin family. Contains 3 ubiquitin-like domains.