Ubiquitin-Proteasome Pathway Assay (CAT#: STEM-MB-0324-WXH)

Introduction

Protein degradation mediated by the ubiquitin proteasome pathway is an important mechanism for the body to regulate the level and function of intracellular proteins. The components responsible for performing this regulatory process include ubiquitin and its initiating enzyme system and proteasome system. The ubiquitin priming enzyme system is responsible for activating ubiquitin and binding it to the protein to be degraded to form a polyubiquitin chain of the target protein, that is, ubiquitination. The proteasome system can recognize and degrade ubiquitinated proteins. In addition, there is another type of deubiquitinated protease that dissociates the ubiquitin chain molecule in the cell to form a reverse regulation.

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Principle Applications Procedure Materials Notes Related Products

Principle

Ubiquitin-proteasome-mediated protein degradation pathways mainly include:
• The activation of ubiquitin: the carboxyl group of the glycine end of ubiquitin is connected to the sulfhydryl group of ubiquitin activating enzyme E1. This step requires ATP as energy to finally form a thioester bond between ubiquitin and ubiquitin activating enzyme E1.
• E1 delivers the activated ubiquitin to the ubiquitin-conjugating enzyme E2 through the esterification process.
• Ubiquitin ligase E3 connects E2-bound ubiquitin to the target protein and releases E2 to form a specific ubiquitinated protein.
• The ubiquitinated protein is recognized and bound by a specific proteasome, and finally the protein is broken down into short peptides or amino acids under the catalysis of proteases.

Applications

To study the regulation mechanism of ubiquitin proteasome signal pathway in disease
To study the effect of each virus on ubiquitin proteasome signaling pathway
To study the effects of drugs or therapies on ubiquitin proteasome signaling pathways

Procedure

• Luminex Multiplex Assay
• Enzyme-linked immunosorbent assay (ELISA)
• Flow cytometry (FACS analysis) technology

Notes

Detectable targets: E1, BID, IAPs, TLR3, SODD, Bad, E2, Bax, PIDD, MYD88, Rac1, TLR4, E3, RIP1, TRAF-2, FADD, APAF-1, BID, FADD, IRF7, Mda-5, NFκB, RIG-1, TRAF3, GAS, IRF9, MEKK1, p38, RIP1, TRAF5, IKK-α, IRS1, MEK6, p50, SLP76, TRAM, IKK-β, IRS2, MSK1, p65, Tak1, TRIF, IPS-1, ISGF3, MSK2, PI3K, TBK1, Vav, IRAK1, ISRE, mTOR, PKR, TLR3

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