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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.

SUBMITTER: Cottrell JA

PROVIDER: S-EPMC4257868 | biostudies-literature | 2015 Jan

REPOSITORIES: biostudies-literature

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Method for measuring lipid mediators, proteins, and messenger RNAs from a single tissue specimen.

Cottrell Jessica A JA Lin Hsuan-Ni HN O'Connor J Patrick JP

Analytical biochemistry 20141020

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 ...[more]

PMID: 25447462

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