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| Product Name | Free Ubiquitin Chain Detection Kit |
| Description |
Capture and detection of unanchored polyubiquitin chains |
| Species Reactivity | Species Independent |
| Platform | Spin Column |
| Sample Types | Cell lysates, Tissue |
| Assay Type | Binding Matrix/Quantitative WB (Western Blot) |
| Utility | Detection kit used to capture, detect, identify and characterise unanchored polyubiquitin chains in samples. |
| Incubation Time | 2 hours |
| Number of Samples | 10 samples |
| Other Resources | Kit Booklet , MSDS |
| Field of Use | Not for use in humans. Not for use in diagnostics or therapeutics. For in vitro research use only. |
Properties
| Storage Temperature | 4ºC | |||||||||||||||
| Shipping Temperature | Blue Ice | |||||||||||||||
| Product Type | Detection Kits | |||||||||||||||
| Assay Overview | The Free Ubiquitin Chain Detection kit facilitates the fast, effective capture and detection of unanchored polyubiquitin chains and free ubiquitin from biological samples. The kit utilises a high capacity binding matrix, specific for the free C-terminus of ubiquitin, together with an easy-to-use purification system for less ‘hands on time’ and superior performance. Allows purification of free ubiquitin and free ubiquitin chains, independent of chain linkage or length. Highly adaptable assay, compatible with samples from a wide range of species and with a broad range of lysis buffers. Analysis by Western blotting or proteomic methods enables identification and analysis of free ubiquitin chains. Kit contains sufficient free ubiquitin chain matrix to perform up to 10 assays. | |||||||||||||||
| Kit Overview |
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| Cite This Product | Free Ubiquitin Chain Detection Kit (StressMarq Biosciences Inc., Victoria BC CANADA, Catalog # SKT-130) |
Biological Description
| Alternative Names | Polyubiquitin B Detection Kit, RPS27A Detection Kit, UBA52 Detection Kit, UBB Detection Kit, UBC Detection Kit, ubiquitin B Detection Kit |
| Research Areas | Apoptosis, Autophagy, Cancer, Cardiovascular System, Cell Signaling, Neurodegeneration, Neuroscience, Post-translational Modifications, Ubiquitination |
| Scientific Background | Ubiquitin typically exerts its fundamental role in the regulation of cellular function through covalent attachment of its C-terminal glycine (Gly76) to lysine residue(s) on substrate proteins (ubiquitination), often in the form of isopeptide linked polyubiquitin chains. Non-substrate linked polyubiquitin chains, termed free or unanchored, have also been identified and been shown to play important roles in a range of cellular processes. Ubiquitin signalling using free ubiquitin chains is propagated via their non-covalent recognition by ubiquitin binding proteins. Processes involving free ubiquitin chains include: secondary messaging in NFkappaB signalling pathways (1,2); stress response (3); regulation of innate immunity (4); regulation of aggresomes (5,6); and as regulators of proteasome activity (7,8). The origins of free ubiquitin chains have yet to be fully elucidated though specific ubiquitin E3 ligases (1,4) and deubiquitinating enzymes (DUBs) (5,9,10) are thought to play a role in their production. As with polyubiquitin modified substrate proteins, the isopeptide linkages that connect the ubiquitin moieties of specific free ubiquitin chains are likely to define their particular function. Examples to date include: free K63-linked polyubiquitin chain activation of TAK1 kinase1 and RIG-I4; chains with linkages alternate to K63 (and K48) in activation of the IKK complex1; and free K48-linked ubiquitin chains in regulation of the 26S proteasome (8). |
| References |
1. Xia, Z.-P. et al. Nature 461, 114–9 (2009). 2. Chen, J. & Chen, Z. J. Curr. Opin. Immunol. 25, 4–12 (2013). 3. Braten, O., Shabek, N., Kravtsova-Ivantsiv, Y. & Ciechanover, A. Biochem. J. 444, 611–7 (2012). 4. Zeng, W. et al. Cell 141, 315–30 (2010). 5. Ouyang, H. et al. J. Biol. Chem. 287, 2317–27 (2012). 6. Hao, R. et al. Mol. Cell 51, 819–28 (2013). 7. Amerik AYu, Swaminathan, S., Krantz, B. a, Wilkinson, K. D. & Hochstrasser, M. EMBO J. 16, 4826–38 (1997). 8. Dayal, S. et al. J. Biol. Chem. 284, 5030–41 (2009). 9. Hanna, J. et al. Cell 127, 99–111 (2006). 10. Zhang, N.-Y., Jacobson, A. D., Macfadden, A. & Liu, C.-W. J. Biol. Chem. 286, 25540–6 (2011). |
Product Images
Western blot analysis of StressXpress® Free Ubiquitin Chain Detection Kit (SKT-130) matrix capture of free ubiquitin and free ubiquitin chains from HEK293T lysate following heat treatment of samples. Ubiquitin species present in lysate sample before (1) and after (2) heat treatment and in elution fraction (3) were detected by ubiquitin polyclonal antibody at 1:1000 dilution.
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