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mTOR Signaling Pathway Luminex Assay (CAT#: STEM-MB-0351-WXH)

Introduction

The mammalian target of rapamycin (mTOR) is a key protein evolutionarily conserved from yeast to man and is essential for life. The mTORC1 complex is made up of mTOR, Raptor, mLST8, and PRAS40, and the mTORC2 complex is composed of mTOR, Rictor, Sin1, and mLST8.
mTOR signaling pathway serves as a master regulator of cell metabolism, growth, proliferation and survival. mTORC1 positively regulates cellular metabolism and ATP production, as well as controlling the synthesis of lipids for membrane generation in proliferating cells and promoting cellular growth by negatively regulating autophagy. Aberrant mTOR signaling is involved in many pathological conditions including cancer, cardiovascular disease, diabetes, obesity, and neurodegeneration. Thus, mTOR inhibitor use in treating solid tumors, rheumatoid arthritis, coronary restenosis, organ transplantation, and other diseases is growing and increasing our knowledge of the role of the mTOR pathway in health, disease, and aging.




Principle

This pathway mainly uses growth factors as the initial signaling molecule required for activation. These growth factors bind to tyrosine kinase receptors in the cell membrane, which activates the pathway. Many of the enzymes involved in this pathway are activated by phosphorylation, when a phosphate group is transferred onto the target enzyme causing it to become active and continue the pathway to other targets.
After growth factors bind to the receptors on the membrane:
• The enzyme PI3 kinase is activated by phosphorylation
• PI3 kinase phosphorylates the substrate PIP2 into PIP3
• PIP3 activates the enzyme Akt, which then activates the protein mTOR
Then, mTOR activates many different transcription factors, which enter the nucleus of the cell and affect the transcription and expression of certain genes involved in cell growth and metabolism.

Applications

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

Procedure

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

Notes

Detectable targets: TNF, R, LRP-5, LRP-6, HSPG, Wnt, IRSI, PIP2, PIP3, PTEN, Ras, Raf, MEK1, MEK2, ERK1, ERK12, RSK, APC, GSK-3beta, RIP1, NIK, IKK, beta, REDD1, p53, MO25, AMPK, LKBI, eEF2, eEF2K, eIF4A, eIF4B, eIF4E, eIF4G, PDCD4, 4EBP1, Maf1, SGK1, AKt, TSC1, TSC2, PDK-1, Rac1, Paxillin, S6, Lipin1, SREBP1, SREBP2, TIFIA, Pol, I, Pol, III

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