Determination of intracellular heat shock protein 70 using a newly developed cell lysate immunometric assay

Rose Njemini, Christian Demanet, Tony Mets

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Heat shock proteins (Hsp) have been associated to several clinical relevant conditions. Currently used methods to determine Hsp 70 possess certain drawbacks. Therefore, we developed a cell lysate immunometric assay (CLIA) for the quantification of intracellular Hsp 70. This CLIA uses a combination of two distinct monoclonal antibodies that recognize different epitopes on the Hsp 70 molecule. A recombinant human Hsp 70 was used as the standard material. The detection range of the CLIA was 4-4000 ng/ml. The intra- and interassay coefficients of variation were, on average, 5% and 12%, respectively. The recovery varied between 81% and 116%. The Hsp 70 levels assayed after serial dilution of cell lysates varied linearly with dilution (between 97% and 120%).

The reliability of the CLIA was assessed by comparison with the values determined by flow cytometric procedure; these two sets of values showed a highly significant correlation (r=0.896, p
Original languageEnglish
Pages (from-to)271-279
Number of pages9
JournalJournal of Immunological Methods
Volume274
Publication statusPublished - 2003

Keywords

  • Heat shock protein 70
  • Cell lysates
  • ELISA
  • Flow cytometry

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