title: Profiling the expression of mitogen-induced T-cell proteins by using multi-membrane dot-blotting.
abstract: 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.
authors.affiliation: 20/20 Gene Systems, Inc., Rockville, MD, USA.
authors.forename: June L
authors.initials: JL
authors.lastname: Traicoff
authors.forename: Mikhail M
authors.initials: MM
authors.lastname: Galperin
authors.forename: Arun
authors.initials: A
authors.lastname: Ramesh
authors.forename: Wendy J
authors.initials: WJ
authors.lastname: Freebern
authors.forename: Cynthia M
authors.initials: CM
authors.lastname: Haggerty
authors.forename: Kevin
authors.initials: K
authors.lastname: Gardner
authors.forename: Vladimir
authors.initials: V
authors.lastname: Knezevic
mesh_terms: D019921:Biotinylation; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism; D015195:Drug Design; D006801:Humans; D019169:Jurkat Cells; D008214:Lymphocytes / Q000378:metabolism; D008934:Mitogens; D010766:Phosphorylation; D040081:Protein Array Analysis / Q000379:methods*; D025941:Protein Interaction Mapping; D020543:Proteome; D015398:Signal Transduction; D013050:Spectrometry, Fluorescence; D013601:T-Lymphocytes / Q000378:metabolism
chemical_list: D008934:Mitogens; D020543:Proteome