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Detection of constitutive and inducible HSP70 proteins in formalin fixed human brain tissue

机译:在福尔马林固定的人脑组织中检测组成型和诱导型HSP70蛋白

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摘要

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.
机译:福尔马林固定和石蜡包埋组织的研究是法医组织学中的常规方法。由于这些样本通常会保存数十年,因此它们为不同的科学问题提供了独特的组织库。过去,使用不同种类的石蜡包埋组织进行了大量研究。然而,众所周知,福尔马林会影响大分子,因此可能会妨碍可靠和可重复的分子实验。本研究的目的是确定福尔马林处理是否对不同的蛋白质检测方法产生负面影响,并另外确定福尔马林的大小。我们将大脑组织样本在福尔马林中孵育了三个月。孵育后,使用免疫组织化学(IHC)和Western印迹分析样品,以特异性检测和定量HSP70超家族成员(热激蛋白)。我们的研究表明,福尔马林固定组织的蛋白质印迹分析根本无法可靠地检测蛋白质,而孵育一个月后仍可以通过IHC进行可重复检测。

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