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Microarrays pp 339-361 | Cite as

Printing Low Density Protein Arrays in Microplates

  • Robert S. Matson
  • Raymond C. Milton
  • Michael C. Cress
  • Tom S. Chan
  • Jang B. Rampal
Part of the Methods in Molecular Biology™ book series (MIMB, volume 381)

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 × 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 × 3 to 5 × 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.

Key Words

Antibodies Biomek cytokines immunoassay microarray microplate 

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Copyright information

© Humana Press Inc., Totowa, NJ 2007

Authors and Affiliations

  • Robert S. Matson
    • 1
  • Raymond C. Milton
    • 1
  • Michael C. Cress
    • 1
  • Tom S. Chan
    • 1
  • Jang B. Rampal
    • 1
  1. 1.Beckman Coulter, Inc.Fullerton

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