Bone protein extraction without demineralization using principles from hydroxyapatite chromatography.

ABSTRACT: Historically, extraction of bone proteins has relied on the use of demineralization to better retrieve proteins from the extracellular matrix; however, demineralization can be a slow process that restricts subsequent analysis of the samples. Here, we developed a novel protein extraction method that does not use demineralization but instead uses a methodology from hydroxyapatite chromatography where high concentrations of ammonium phosphate and ammonium bicarbonate are used to extract bone proteins. We report that this method has a higher yield than those with previously published small-scale extant bone extractions, with and without demineralization. Furthermore, after digestion with trypsin and subsequent high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) analysi

SUBMITTER: Cleland TP 

PROVIDER: S-EPMC4314460 | biostudies-literature | 2015 Mar

REPOSITORIES: biostudies-literature