UbiQapture™-Q kit by Enzo Life Sciences

For isolation and enrichment of ubiquitinylated proteins.The UbiQapture™-Q Kit is an efficient tool for the selective isolation of ubiquitinylated proteins. The Kit facilitates the isolation of both mono- and poly-ubiquitinylated proteins (independent of lysine residue chain linkage) from cell extracts, tissue lysates and in vitro assay solutions through use of a high-binding affinity matrix. Captured proteins can be analyzed by Western blotting using the highly sensitive ubiquitin-conjugate specific antibody provided, using antibodies to specific proteins of interest, or eluted from the matrix for subsequent biochemical characterization. The UbiQapture™-Q matrix supplied with the Kit has superior binding characteristics compared to other commercially available matrices and is compatible with a wide range of lysate buffers and cell/tissue samples from a variety of species.Suggested uses: Isolation and detection of ubiquitinylated protein conjugates from a specific cell/tissue lysate, Capture and analysis of specific ubiquitinylated protein conjugates of interest from particular cell/tissue lysates, Release of free proteins in their active/native form by cleavage of ubiquitin/ubiquitin chains from the UbiQapture™-Q matrix using a deubiquitinylating enzyme, Release of ubiquitinylated proteins in their active/native form by elution from the UbiQapture™-Q matrix using high salt buffer.

1. Isolation and detection of the full range of ubiquitinylated protein conjugates (mono-/multi-/poly-ubiquitin modified, lysine linkage independent) from specific cell/tissue lysates of interest, especially low abundance (regulatory) proteins involved in the ubiquitin-proteasome pathway.
2. Capture and analysis of specific ubiquitinylated protein conjugates of interest from particular cell/tissue lysates.
3. Purification/pull down of ubiquitinylated proteins from cell free in vitro assays; for example to isolate Ub-p53 species from ubiquitinylation assays mediated by HeLa S100 lysate (Prod. No. BML-SW8750).
4. Release of free proteins in their active/native form by cleavage of ubiquitin/ubiquitin chains from the UbiQapture-Q matrix using deubiquitinylating enzymes such as USP2 (Prod. No. BML-UW9850).
5. Release of ubiquitinylated proteins in their active/native form by elution from the UbiQapture-Q matrix using, for example, high salt buffer.

Note: Protocol provided covers applications 1-3. Assay set-up can be readily modified for alternative applications by inclusion, omission or
substitution of specific components.
Kit is not suitable for use with purified proteins