title: Analysis of Gene and Protein Expression in Atherosclerotic Mouse Aorta by Western Blot and Quantitative Real-Time PCR.
abstract: Atherosclerosis involves changes in gene and protein expression patterns in affected arteries. Quantification of these alterations is essential for understanding the molecular mechanisms underlying this pathology. Western blot and real-time PCR-used to quantify protein and messenger RNA levels, respectively-are invaluable molecular biology tools, particularly when material is limited. The availability of many genetically modified mouse models of atherosclerosis makes the mouse aorta an ideal tissue in which to carry out these expression pattern analyses. In this chapter, protocols are presented for mRNA and protein extraction from mouse aorta and for the accurate quantification of mRNA expression by RT-PCR and of proteins by western blot.
authors.affiliation: Laboratory of Molecular and Genetic Cardiovascular Pathophysiology, Vascular Biology Program, Centro Nacional de Investigaciones Cardiovasculares Carlos III (CNIC), Melchor Fernández Almagro 3, 28029, Madrid, Spain. jrivera@cnic.es.
authors.forename: José
authors.initials: J
authors.lastname: Rivera-Torres
mesh_terms: D000818:Animals; D001011:Aorta / Q000378:metabolism*; D001018:Aortic Diseases / Q000235:genetics* / Q000378:metabolism*; D050197:Atherosclerosis / Q000235:genetics* / Q000378:metabolism*; D015153:Blotting, Western / Q000379:methods*; D004195:Disease Models, Animal; D005786:Gene Expression Regulation; D005819:Genetic Markers; D051379:Mice; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors
chemical_list: D005819:Genetic Markers; D012333:RNA, Messenger
keywords: Protein; RNA; Real-time PCR; Reverse transcription; SYBR Green; Western blot; cDNA