Detection of constitutive and inducible HSP70 proteins in formalin fixed human brain tissue.
The investigation of formalin fixed and paraffin embedded tissue is a routine method in forensic histology. Since these samples are usually stored for decades they provide a unique tissue bank for different scientific issues. In the past, numerous studies were conducted using different kinds of paraffin embedded tissues. However, it is well known that formalin affects macromolecules and thus might hamper reliable and reproducible molecular experiments. The aim of this study was to find out if the treatment with formalin has a negative effect on different protein detection methods and additionally to define the dimension of those possible deleterious effects. We incubated brain tissue samples in formalin for up to three months. After incubation, the samples were analyzed using immunohistochemistry (IHC) and Western blotting to specifically detect and quantify members of the HSP70 superfamily (heat shock proteins). Our study shows that the Western blot analysis of formalin fixed tissues does not allow a reliable detection of proteins at all, while a reproducible detection by IHC was still possible after one month of incubation.
Institute of Legal Medicine, University Hospital Schleswig-Holstein, Campus Kiel, Germany. Electronic address: Andrea.Preusse-Prange@uksh.de.
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Preusse-Prange
Institute of Legal Medicine, University Hospital Schleswig-Holstein, Campus Kiel, Germany.
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Modrow
Institute of Legal Medicine, University Hospital Schleswig-Holstein, Campus Kiel, Germany.
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Schwark
Institute of Legal Medicine, University Hospital Schleswig-Holstein, Campus Kiel, Germany.
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von Wurmb-Schwark
D000368:Aged; D015153:Blotting, Western; D002531:Cerebellum / Q000378:metabolism* / Q000473:pathology; D054022:Cerebrum / Q000378:metabolism* / Q000473:pathology; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005404:Fixatives; D005557:Formaldehyde; D050883:HSC70 Heat-Shock Proteins / Q000378:metabolism*; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008875:Middle Aged; D013048:Specimen Handling; D013997:Time Factors
D005404:Fixatives; D050883:HSC70 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; C490786:HSPA1A protein, human; C494371:HSPA8 protein, human; D005557:Formaldehyde
Formalin; Heat shock protein; Immunohistochemistry; Western blot