PI3K-Akt Pathway Luminex Multiplex Assay

PI3K-Akt signaling pathway

Based on Luminex technology platform, Creative Proteomics provides analysis services for key targets of PI3K-Akt signaling pathway.

PI3K-Akt Pathway is an intracellular signal transduction pathway that promotes metabolism, proliferation, cell survival, growth and angiogenesis in response to extracellular signals. This is mediated through serine and/or threonine phosphorylation of a range of downstream substrates. Key proteins involved are phosphatidylinositol 3-kinase (PI3K) and Akt/Protein Kinase B.

The key molecules involved in this signaling pathway are receptor tyrosine kinase (RTKs), phosphatidylinositol 3-kinase (PI3K), phosphatidylinositol-4,5-bisphosphate (PIP2), phosphatidylinositol-3,4,5-bisphosphate (PIP3) and AKT/protein kinase B.

The PI3K-Akt pathway has many downstream effects and must be carefully regulated. Negatively regulation of PI3K-AKT pathway can be achieve at to target: the PIP3 level and the inactivation of AKT protein. Phosphatase and tensin homolog (PTEN) is a main down regulation protein which can converting PIP3 into PIP2. Protein phosphatase 2A (PP2A), which dephosphorylates Akt at Thr308 and phosphatase PHLPP dephosphorylates Akt at Ser473 are also two negative regulation proteins. Addition to these regulation protein, the pathway itself also have feedback mechanisms: Transcription factor NF-κB, activated by Akt, regulates peroxisome proliferator-activated receptor delta (PPARβ/δ) agonists and tumor necrosis factor α (TNFα), which in turn repress PTEN expression as a positive feedback; Negative feedback loop is functioned by mTORC1 and S6K1 activation. S6K1 is able to phosphorylate IRS-1 at multiple serine residues, preventing binding to RTKs, resulting the suppression of PI3K activation.

As the general regulation direction of PI3K-AKT pathway is to stimulate the cell growth and proliferation. The over activation of this signaling pathway can over stimulate cell and resulting abnormal cell proliferation, that is oncogenesis. Abnormalities in the PI3K pathway are common in cancer and have a role in neoplastic transformation . PI3K itself is a frequent target of mutational activation. The most frequent genetic aberrations in breast cancer are somatic missense mutations in the PIK3CA gene encoding p110α. There are two kinds of mutations of PIK3CA gene, abnormalities of gene copy number and point mutation that resulting the persistently activation of PI3K without the RTKs stimulation. 80% of the point mutation of PI3K are E542K, E545K and H1047R mutation. The patients with these point mutation are insensitive to the targeting anticancer drugs of EGFR and Her2 inhibitor.

Our detectable targets:

Technology platform

We provide Luminex technology for PI3K-Akt signaling pathway analysis.

Luminex technology is a multifunctional liquid phase analysis platform developed on the basis of colored microspheres, laser technology, applied fluidics and high-speed digital signal processing technology. The core is to encode polypropylene microspheres or magnetic microspheres with fluorescent dyes. By adjusting the different ratios of the two fluorescent dyes, up to 100 microspheres with different fluorescence spectra can be obtained. Each kind of microspheres is covalently cross-linked. Capture antibodies against specific antigens.

The PI3K-Akt pathway is an intracellular signal transduction pathway that plays a major role in promoting the metabolism, proliferation, cell survival, growth and angiogenesis of extracellular signals.

In addition to Luminex Multiplex Assay, Enzyme-linked immunosorbent assay (ELISA), Flow cytometry (FACS analysis) technology can also be provided to meet other customer needs.

Advantages of PI3K-Akt signaling pathway detection:

• High flexibility: specific probes, antigens or antibodies can be connected to the microspheres to meet the needs of different customers.
• High sensitivity: the detection limit can reach 0.01pg/ml.
• Good repeatability: similar homogeneous reaction mode, each indicator has 1000-5000 reaction units, and the median value of 100 analysis is taken.

Application of our service:

• To study the effect of each virus on PI3K-Akt signaling pathway
• To study the regulation mechanism of PI3K-Akt signal pathway in disease
• To study the effects of drugs or therapies on PI3K-Akt signaling pathways

Creative Proteomics has developed a signal pathway target detection platform. We are not limited to providing PI3K-Akt signal path detection services, but can also provide other signal path detection services. If you want to detect other targets, please contact us and we will customize the service for you. Look forward to working with you.

References:

1. Hennessy B T, Smith D L, et al. Exploiting the PI3K/AKT pathway for cancer drug discovery. Nature reviews Drug discovery, 2019, 4(12): 988-1004.
2. Osaki M, Oshimura M, et al. PI3K-Akt pathway: its functions and alterations in human cancer. Apoptosis, 2020, 9(6): 667-676.