Comparison of 2 Luminex-based Multiplexed Protein Assays for Quantifying Microglia Activation and Inflammatory Proteins

Abstract

The complex injury cascades associated with mild traumatic-brain injury partly involve inflammation in the brain that is triggered by the release of chemokines and cytokines from the brains resident immune cells, microglia. The technical ability to quantify these small molecules from biological samples is critical toward new technology that limits the damage following a concussive event. This projects objective was to test the feasibility of using multiplexed protein assays based on Luminex microspheres to fulfil the requirement to quantify inflammatory proteins from biological samples derived from rats. Two vendors, R and D Systems and EMD Millipore, market antibodies conjugated with Luminex fluorescently labeled microspheres that can be used to quantify inflammatory biomarkers directly from samples of plasma, cell cultures, tissue homogenates, cerebral spinal fluid, and other tissues. The single kit from R and D Systems removed many sources of human error and provides a diluent to generate a single standard curve for a variety of sample types. EMD Millipore has many more products available to detect a wide range of analytes in rat samples, but these products are separated into different kits. Without optimization, the products from both vendors were only capable of relative quantification of fluorescence instead of true quantification of concentration.

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Document Details

Document Type
Technical Report
Publication Date
Apr 01, 2016
Accession Number
AD1007075

Entities

People

  • Brandon Harvey
  • Emily Wires
  • Keith W. Whitaker
  • Lee Campbell

Organizations

  • United States Army Research Laboratory

Tags

Communities of Interest

  • Biomedical

DTIC Thesaurus Topics

  • Antibodies
  • Biological Sciences
  • Blood
  • Blood Plasma
  • Brain Injuries
  • Cell Physiological Processes
  • Cells
  • Culture Techniques
  • Cytokines
  • Department Of Defense
  • Drug Abuse
  • Fluorescence
  • Medical Personnel
  • Microspheres
  • Optical Properties
  • Proteins
  • Standards

Fields of Study

  • Biology

Readers

  • Immunology and Pathology
  • Molecular and Cellular Biochemistry
  • Oncology and Biomarker-Based Cancer Detection.