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Interactions Between Signaling Pathways

Interactions Between Signaling Pathways

Signaling cascades usually lack linearity. Multiple pathways interact and regulate one another, allowing cells to integrate and respond to diverse environmental stimuli.
Convergence and divergence, and cross-talk between signaling pathways
Two distinct signaling pathways can converge on a single functional unit, which may either be a single protein or a complex of proteins. The response is either functionally distinct or synergistic between the two pathways but different from the response...

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Updated: Nov 28, 2025

Assessment of Resistance to Tyrosine Kinase Inhibitors by an Interrogation of Signal Transduction Pathways by Antibody Arrays

Assessment of Resistance to Tyrosine Kinase Inhibitors by an Interrogation of Signal Transduction Pathways by Antibody Arrays

Published on: September 19, 2018

Analyzing Signaling Pathways Using Antibody Arrays.

Hao Tang¹, Chaohui Duan², Zhizhou Kuang^3,4

¹RayBiotech Life, Peachtree Corners, GA, USA. Hao@raybiotech.com.

Methods in Molecular Biology (Clifton, N.J.)

|November 25, 2020

Summary

This summary is machine-generated.

We developed a high-throughput antibody array to profile cell signaling pathways. This technology efficiently analyzes protein phosphorylation changes induced by drugs, aiding disease research and drug development.

Keywords:

Antibody array Apoptosis MAPK Signaling pathway

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Area of Science:

Molecular Biology
Cell Signaling
Biotechnology

Background:

Cell signaling pathways are crucial for homeostasis and disease.
Traditional methods for studying these pathways are challenging.
Understanding signaling networks can improve disease pathology insights and drug development.

Purpose of the Study:

To develop a high-throughput antibody array for simultaneous detection of multiple signaling pathway proteins.
To investigate the effects of anti-cancer drugs on protein phosphorylation.
To validate the efficiency and accuracy of the antibody array technology.

Main Methods:

Developed a sandwich-based antibody array on nitrocellulose or glass slides.
Spotted arrays with antibodies targeting key proteins in RTK, EGFR, MAPK, AKT, apoptosis, TGFb, JAK/STAT, NFkB, and insulin receptor pathways.
Applied arrays to analyze protein phosphorylation changes in Jurkat and HeLa cells treated with camptothecin and PMA, respectively, validating with Western blotting.

Main Results:

Camptothecin treatment induced DNA damage and apoptosis via ATM, Chk2, caspase 3, and PARP in Jurkat cells.
Phorbol 12-myristate 13-acetate (PMA) activated the MAPK pathway, including ERK, CREB, and RSK1, in HeLa cells.
Array data were consistent with previous studies and validated by Western blotting.

Conclusions:

Pathway antibody arrays offer a rapid, efficient, and multiplexed method for profiling phosphorylated proteins.
This technology aids in understanding cell signaling dynamics in response to stimuli.
The developed arrays are valuable tools for disease research and drug discovery.