Profiling the expression of mitogen-induced T-cell proteins by using multi-membrane dot-blotting

High throughput technologies are standard methods for analysis of the proteome. Multi-layer multi-well plate dot-blotting system (MLDot) technology is a high-throughput dot blotting system that provides a simple, cost-effective approach for protein expression profiling in multiple samples. In contra...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 2004-10, Vol.323 (1), p.355-360
Main Authors: Traicoff, June L., Galperin, Mikhail M., Ramesh, Arun, Freebern, Wendy J., Haggerty, Cynthia M., Gardner, Kevin, Knezevic, Vladimir
Format: Article
Language:English
Subjects:
Biotinylation
Blotting, Western
Cell Membrane - metabolism
Cell signaling
Drug Design
Drug discovery
Humans
Jurkat Cells
Lymphocytes - metabolism
Mitogens
Multiplex
Neoplasia
Phosphorylation
Protein Array Analysis - methods
Protein expression
Protein Interaction Mapping
Proteome
Signal Transduction
Spectrometry, Fluorescence
T-Lymphocytes - metabolism
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Summary:High throughput technologies are standard methods for analysis of the proteome. Multi-layer multi-well plate dot-blotting system (MLDot) technology is a high-throughput dot blotting system that provides a simple, cost-effective approach for protein expression profiling in multiple samples. In contrast to traditional dot blot, MLDot uses a layered stack of thin, sieve-like membranes in place of a single nitrocellulose membrane. Therefore, up to 10 membranes can be prepared from the samples arrayed in a single 96-well plate. We describe the ability of MLDot to detect the predicted changes in protein expression following multiple mitogen treatment of T-cells. We compare the levels of the phopshorylated forms of CREB, Jun, and Akt in Jurkat T-cells as detected by MLDot to those measured by a gel-based assay. We also describe the ability of MLDot to detect differences in the levels of phosphorylated Akt in Jurkat cells as compared to primary lymphocytes.
ISSN:0006-291X
1090-2104
DOI:10.1016/j.bbrc.2004.08.093