title: Method for measuring lipid mediators, proteins, and messenger RNAs from a single tissue specimen.
abstract: This article describes a new method for extracting RNA, protein, and lipid mediators from a single tissue specimen. Specifically, mouse bone fracture callus specimens were extracted into a single solution that was processed using three different procedures to measure messenger RNA (mRNA) levels by reverse transcription-quantitative polymerase chain reaction (RTqPCR), cytokines and growth factors using an xMAP method, and lipid mediators by liquid chromatography-tandem mass spectrometry (LC-MS/MS). This method has several advantages because it decreases the number of animals necessary for experimentation, allows division of the sample from a homogeneous mixture that reduces sample variability, and uses a solution that protects the integrity of the macromolecules during storage.
authors.affiliation: Department of Biochemistry and Molecular Biology, New Jersey Medical School, Rutgers, The State University of New Jersey, Newark, NJ 07103, USA.
authors.forename: Jessica A
authors.initials: JA
authors.lastname: Cottrell
authors.affiliation: Department of Biochemistry and Molecular Biology, Graduate School of Biomedical Sciences, Rutgers, The State University of New Jersey, Newark, NJ 07103, USA.
authors.forename: Hsuan-Ni
authors.initials: HN
authors.lastname: Lin
authors.affiliation: Department of Biochemistry and Molecular Biology, New Jersey Medical School, Rutgers, The State University of New Jersey, Newark, NJ 07103, USA; Department of Biochemistry and Molecular Biology, Graduate School of Biomedical Sciences, Rutgers, The State University of New Jersey, Newark, NJ 07103, USA. Electronic address: oconnojp@njms.rutgers.edu.
authors.forename: J Patrick
authors.initials: JP
authors.lastname: O'Connor
mesh_terms: D000818:Animals; D002146:Bony Callus / Q000378:metabolism / Q000473:pathology; D002851:Chromatography, High Pressure Liquid; D016207:Cytokines / Q000032:analysis; D050723:Fractures, Bone / Q000378:metabolism* / Q000473:pathology; D036341:Intercellular Signaling Peptides and Proteins / Q000032:analysis*; D008055:Lipids / Q000032:analysis* / Q000302:isolation & purification; D051379:Mice; D012333:RNA, Messenger / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D052616:Solid Phase Extraction; D053719:Tandem Mass Spectrometry
chemical_list: D016207:Cytokines; D036341:Intercellular Signaling Peptides and Proteins; D008055:Lipids; D012333:RNA, Messenger
keywords: Bone regeneration; Fracture healing; Lipid mediator; Metabolomics