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A novel ELISA for mouse activated protein C in plasma

Academic Article
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Overview

authors

  • Fernandez, J. A.
  • Lentz, S. R.
  • Dwyre, D. M.
  • Griffin, John

publication date

  • July 2006

journal

  • Journal of Immunological Methods  Journal

abstract

  • The Protein C pathway plays a crucial role in the regulation of thrombosis and inflammation. One of the tools that researchers presently use to elucidate mechanisms of action of activated protein C (APC) is the use of transgenic or gene deletion murine models. To correlate observations in these murine models with the APC levels, there is a need for a sensitive and specific assay for circulating murine APC in plasma. We developed an immunological assay to measure the physiological and pharmacologic levels of circulating murine APC. The sandwich ELISA uses an anti-murine anti-protein C antibody capture antibody and human protein C inhibitor (PCI) as a detection reagent, taking advantage of the facts that the mouse lacks plasma PCI and that human PCI forms a 1/1 stable complex with mouse APC. The amount of complex APC:PCI is detected with an anti-human PCI monoclonal antibody. The assay shows improved sensitivity versus enzyme immunocapture assays commonly used to detect human APC and considerably reduces the processing time.

subject areas

  • Animals
  • Antibody Specificity
  • Blood Coagulation
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Protein C
  • Protein C Inhibitor
  • Rabbits
  • Sensitivity and Specificity
  • Thrombin
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Research

keywords

  • ELISA
  • activated protein C
  • mouse
  • protein C
  • protein C inhibitor
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Identity

International Standard Serial Number (ISSN)

  • 0022-1759

Digital Object Identifier (DOI)

  • 10.1016/j.jim.2006.05.004

PubMed ID

  • 16828789
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Additional Document Info

start page

  • 174

end page

  • 181

volume

  • 314

issue

  • 1-2

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