Analysis Note

ControlLys63 polyubiquitin chains.

Application

Research CategorySignaling

Anti-Ubiquitin Antibody, Lys63-Specific, clone Apu3 is a high quality Rabbit Monoclonal Antibody for the detection of Ubiquitin and has been validated in Inhibition Assay (PIA), Immunocytochemistry (ICC), Immunoprecipitation (IP), Immunohistochemistry (IHC) and Western Blotting (WB).

Research Sub CategoryUbiquitin & Ubiquitin Metabolism

Immmunoprecipitation:
Rabbit monoclonal anti-ubiquitin (Lys-63-specific), clone Apu3, precipitated ubiquitin-containing proteins from HeLa cell lysates

Immunohistochemistry:
Ubiquitin (Lys63-specific) staining of colorectal cancer tissue sections is restricted to tumor cells.

Flow cytometry:
Flow cytometry analysis of Jurkat cells using Anti-ubiquitin Lys63-specific (#05-1308) (blue) and rabbit IgG control (red).

Inhibition Assay:
Preincubation of anti-ubiquitin, Lys63-specific, with K63-multiubiquitin chains inhibited binding to HeLa cell lysate in Western blots.

Immunocytochemistry:
Ubiquitin (Lys63-specific) staining of colorectal cancer tissue sections is restricted to tumor cells. Positive staining was also detected in mouse, rat, and rabbit brain sections.

Disclaimer

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.

General description

Polyubiquitin chains linked through the Lys63 residue of ubiquitin (Ub) are associated with nonproteolytic functions such as signal transduction. Lys63-linked polyUb chains connect components of NFκB signaling in a highly regulated manner, and genetic evidence indicates the involvement of Lys63-linked Ub chains in stress response and DNA repair. Research in yeast has suggested Lys63-linked Ub chains stimulate endocytosis.

In contrast, polyubiquitin chains linked through the Lys48 residue of ubiquitin are most commonly associated with proteins targeted for proteosomal degradation.

DI-UBIQUITIN PROTEIN AVAILABLE!
Di-Ubiquitin (Lys63-linked) Recombinant Protein (17-10388)

Immunogen

Immunogen used was human di-ubiquitin linked at Lys63.

Epitope: Lys63 ubiquitination site

Physical form

Protein A purified recombinant rabbit monoclonal antibody that consists of a humanized fAb fragment fused to a rabbit Fc tail produced in a CHO recombinant antibody system. Supplied in 0.1M Tris-Glycine (pH7.4) 150 mM NaCl, 0.05% sodium azide.

Protein A purified

Format: Purified

Quality

Western Blot Analysis:
A 1:1000 dilution of this antibody detected Lys-63 ubiquitin chains by Western blotting, but did not detect free ubiquitin or Lys48 polyubiquitin chains.

Specificity

Based on sequence homology, broad species cross-reactivity is expected.

Recognizes ubiquitin linked at Lys63. Does not recognize ubiquitin linked at Lys48 or free ubiquitin.

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.
For maximum recovery of product, centrifuge the vial prior to removing the cap.

Target description

varies