Polyubiquitin B Detection Kit, RPS27A Detection Kit, UBA52 Detection Kit, UBB Detection Kit, UBC Detection Kit, ubiquitin B Detection Kit
Categories
Detection Kits
Description
Capture and detection of ubiquitin binding proteins
Field Of Use
Not for use in humans. Not for use in diagnostics or therapeutics. For in vitro research use only.
Gross Weight (kg)
2
Net Weight (g)
300
Reactivity
Species independent
Scientific Background
The covalent attachment of ubiquitin to proteins (ubiquitination) plays a fundamental role in the regulation of cellular function through biological events including cell cycle, differentiation, immune responses, DNA repair, chromatin structure, transcription, signal transduction, endocytosis, apoptosis and degradation by the proteasome, autophagy and lysosome systems. As such ubiquitin signalling and the processes it mediates are essential for the normal functioning of cells and its dysfunction has been implicated in wide range of diseases including cancer, neurodegeneration, cardiovascular and metabolic disorders (1,2). The type of ubiquitin modification, (monoubiquitin, multiubiquitin, polyubiquitin), substrate protein lysine residue(s) modified and, in the case of polyubiquitination, the chain length and lysine linkage type control the function and fate of ubiquitinated proteins. In addition all ubiquitin mediated pathways also utilise specific ubiquitin receptors to facilitate their regulation (3,4). Ubiquitination is achieved through three enzymatic steps. In an ATP-dependent process, the ubiquitin E1 activating enzyme catalyses the formation of a reactive thioester bond with ubiquitin, followed by its subsequent transfer to the active site cysteine of a ubiquitin E2 carrier protein. The selectivity of the ubiquitin cascade for a particular substrate protein relies on the interaction between the E2 conjugating enzyme (of which a cell contains relatively few) and an ubiquitin E3 ligase , of which over 600 have been identified to date. The specific E2-E3 pair required for ubiquitination of a particular substrate protein in vivo may also control the type, point and length / linkage (polyubiquitin) of the ubiquitin modification (5).
Size
800 µl ; 20 tubes ; 20 tubes
Adr Code
Non-hazardous
Assay Overview
The Ubiquitin Interact Detection kit facilitates the selective capture and detection of ubiquitin binding and associated proteins from cell lysates and tissue extracts. The kit utilises a high capacity, high specificity ubiquitin matrix together with an easy-to-use purification system for efficient isolation of ubiquitin binding proteins with minimal non-specific binding. Allows purification of mono- and poly-ubiquitin binding proteins, independent of chain linkage or length. Analysis by Western blotting or proteomic methods enables identification and assessment of captured proteins. Kit contains sufficient ubiquitin interact matrix to perform up to 20 binding assays.
Detection kit used to capture, detect, identify and characterise isolated ubiquitin binding proteins by Western blotting or proteomic analysis in samples.
Reference
1. Fulda, S., Rajalingam, K. & Dikic, I. EMBO molecular medicine 4, 545–56 (2012). 2. Shaid, S., Brandts, C. H., Serve, H. & Dikic, I. Cell death and differentiation 20, 21–30 (2013). 3. Husnjak, K. & Dikic, I. Annual review of biochemistry 81, 291–322 (2012). 4. Komander, D. & Rape, M. Annual Review of Biochemistry 81, 203–229 (2012). 5. Spasser, L. & Brik, A. Angewandte Chemie (International ed. in English) 51, 6840–62 (2012).