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Stock availability
In Stock
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Kit s description
Capture and detection of ubiquitin binding proteins
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Protein target
Ubiquitin Interact
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Scientific context
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).
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Primary research area
Cell Signaling, Post-Translational Modifications, Ubiquitination, Epigenetics, Cancer, Apoptosis, Autophagy, Neuroscience, Neurodegeneration, Cardiovascular System
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Category
Assay Detection Kits
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Other name
Polyubiquitin B Detection Kit, RPS27A Detection Kit, UBA52 Detection Kit, UBB Detection Kit, UBC Detection Kit, ubiquitin B Detection Kit
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Brand name
StressXpress®
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Detection system
Detection
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Assay format
Binding Matrix/Quantitative WB (Western Blot)
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Main intended usage
Detection kit used to capture, detect, identify and characterise isolated ubiquitin binding proteins by Western blotting or proteomic analysis in samples.
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Brief protocol
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.
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Kit contents
StressXpress® ubiquitin interact matrix (50% slurry), StressXpress® columns, StressXpress® collection tubes
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Assay precision
Contact us
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Assay cross reactivity
Species Independent
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Samples to be used with this kit
Cell Lysates, Tissue
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Maximum samples to be used with this kit
20 samples
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Assay duration
1 hour
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Kit s sensitivity
See product datasheet or feel free to contact us
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Assay detection limit
Contact us
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Storage recommendations
4°C
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Shipping recommendations
Blue Ice
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Bibliography
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).
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Release date
1-Jun-2014
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PubMed number
Refer to PubMed
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Tested applications
No data available / currently testing
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Tested reactivity
currently testing
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Resources available upon request
Kit Booklet, MSDS
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Product image link
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Product image legend
AMC Standard Curve for 20S Proteasome Activity Kit. Representative plot of fluorescence measurements (RFU) of serial dilutions of AMC Standard at recommended concentrations. | Continuous kinetic enzyme activity assay showing the time course of control 20S proteasome activity with Suc-LLVY-AMC substrate using the StressXpress® 20S Proteasome Activity Kit. 20S proteasome (~2.5nM) was incubated with 100µM Suc-LLVY-AMC in proteasome assay buffer for 30 minutes at room temperature alongside a substrate only (blank) control reaction. Fluorescence measurements (RFU) were taken at 30 second intervals and plotted vs. time. Graph of the AMC Typical Standard Curve for the 20S Proteasome Activity Kit StressXpress - SKT-133 | Enzyme Activity Graph of the Continuous Kinetic Assay for the 20S Proteasome Activity Kit StressXpress - SKT-133
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Warnings
Non-hazardous
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Country of production
Canada
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Total weight kg
2
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Net weight g
300
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Description
The detections of the targets with this kit is a type of test that can be performed on any target containing biological samples after clean up of interfering agents. The assay must be performed following the protocol.
