Related Experiment Video
Updated: Aug 8, 2026
Colorectal Cancer Cell Surface Protein Profiling Using an Antibody Microarray and Fluorescence Multiplexing
Published on: September 25, 2011
Panorama Ab Microarray Cell Signaling kit: a unique tool for protein expression analysis
Eliezer Kopf1, Dalia Shnitzer, Dorit Zharhary
1Sigma-Aldrich Israel, Rehovot, Israel.
Insights
This study introduces the Panorama Antibody Microarray, a new tool for analyzing cell signaling proteins. It accurately detected protein changes in differentiating cells, with results confirmed by immunoblotting.
Area of Science:
- Proteomics
- Cell Biology
- Biotechnology
Background:
- Antibody arrays offer a powerful method for high-throughput protein analysis.
- Understanding protein level changes is crucial for studying cellular responses to stimuli.
Purpose of the Study:
- To introduce and evaluate the Panorama Antibody Microarray system for cell signaling analysis.
- To assess the array's performance in detecting protein level changes in differentiating cells.
Main Methods:
- Development of a novel antibody array (Panorama Ab Microarray) with 224 antibodies on nitrocellulose slides.
- Optimization of slide post-spotting treatment for improved performance and stability.
- Application of the array to analyze protein expression changes in F9 embryonic carcinoma cells stimulated with all-trans retinoic acid.
Main Results:
- The Panorama Ab Microarray can detect protein levels as low as nanograms per mL.
- The array identified both up- and down-regulated proteins, including cell cycle regulators and kinases, during F9 cell differentiation.
- Results for over ten protein targets were validated using immunoblotting.
Conclusions:
- The Panorama Ab Microarray is a sensitive and reliable tool for mass analysis of cell signaling proteins.
- The system effectively captures dynamic changes in protein expression during cellular differentiation.
- The array's performance is influenced by post-spotting slide treatment, highlighting the importance of optimization.
Abstract:
Antibody arrays are a promising new tool for mass analysis of protein level changes in cells responding to different stimuli. Here we describe a novel antibody array system called Panorama Ab Microarray Cell Signaling, that contains 224 antibodies spotted on FAST nitrocellulose-coated slides that can detect protein levels as low as a few nanograms per mL. The antibodies spotted are specific for proteins important in various areas of cell signaling such as phosphorylation, cell cycle, apoptosis, nuclear signaling and cytoskeleton proteins. Furthermore, for some of the protein targes, the Panorama Ab Microarray can detect phosphorylated and nonphosphorylated forms of the traget protein. We found that treatment of the slides post-spotting is important for the array performance (ratio of signal to background) and its stability. Panorama Ab Microarray was used to analyze changes in protein expression in F9 embryonic carcinoma cells stimulated to differentiate by all-trans retinoic acid. We found that the level of several proteins, among them cell cycle regulators and kineases, was either up- or down-regulated. For more than ten protein targets, the results obtained by the Panorama Ab Microarray were confirmed by immunoblotting.
