Printing Low Density Protein Arrays in Microplates
2007
Journal Article
Authors:
Matson, R.;
Milton, R.;
Cress, M.;
Chan, T.;
Rampal, J.
Secondary:
Methods Mol Biol
Volume:
381
Pagination:
339-361
URL:
http://www.ncbi.nlm.nih.gov/pubmed/17984528
Keywords:
Animals Antibodies; Cytokines; Enzyme-Linked Immunosorbent Assay; Humans; Mice; Protein Array Analysis; Software
Abstract:
Here, we provide methods for the creation of protein microarrays in microplates. The microplate consists of 96 wells with each well capable of holding a protein microarray at a spot density of up to 400 (20 x 20) individual elements. Arrays of capture monoclonal antibodies, corresponding to specific interleukins, were printed onto the bottom of the wells which had been surface activated for covalent attachment. A Biomek 2000 laboratory automation workstation (Beckman Coulter, Inc., Fullerton, CA) equipped with a high-density replicating tool was used for printing low density 3 x 3 to 5 x 5 arrays. For higher density arrays, a microarrayer system (Cartesian PS7200, Genomic Solutions, Inc., Ann Arbor, MI) was employed. Multiple antigens were simultaneously analyzed without detectable cross-reactivity associated with capture antibody or secondary antibody interactions. Detection of interleukin antigens, spiked into cell culture media containing 10% fetal calf serum, was specific and sensitive.
