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Highly reliable quantification of proteins such as members of the HSP70 superfamily based on the grey scale index via immune detection stained bands on a Western blot
Authors:Jan-Hendrik Modrow  Andrea Preusse-Prange  Patrick Meyer  Melanie Harder  Thorsten Schwark  Nicole von Wurmb-Schwark
Institution:Institute of Legal Medicine Kiel, Arnold-Heller-Strasse 12, D-24105 Kiel, Germany.
Abstract:The HSP70 superfamily is a reliable biomarker for hyperthermia, hypothermia and hypoxia. The Enzyme-linked Immunosorbent Assay (ELISA) respectively immunohistochemically staining methods are the typically used techniques for the quantification of those proteins. As the costs for reagents and devices as well as the work schedule of these methods are immense it was the goal of our study to develop an easy and reliable method to quantify the concentration of specific proteins. We established a procedure to measure the relative concentration of proteins fixed on ROTI(?) PVDF membranes via Western blot, calculating the relative protein concentration in dependency to the grey scale index of the normalized and digitalized pictures of the bands on the blots.
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