ABSTRACT
The sequencing of the complete genome for many organisms, including man, has opened the door to the systematic understanding of how complex structures such as the brain integrate and function, not only in health but also in disease. This blueprint, however, means that the piecemeal analysis regimes of the past are being rapidly superseded by new methods that analyse not just tens of genes or proteins at any one time, but thousands, if not the entire repertoire of a cell population or tissue under investigation. Using the most appropriate method of analysis to maximise the available data therefore becomes vital if a complete picture is to be obtained of how a system or individual cell is affected by a treatment or disease. This review examines what methods are currently available for the large scale analysis of gene and protein expression, and what are their limitations.
Subject(s)
Brain Diseases/genetics , Brain/physiopathology , Dementia/genetics , Genomics/methods , Mental Disorders/genetics , Proteomics/methods , Brain Diseases/physiopathology , Brain Diseases/therapy , Dementia/physiopathology , Dementia/therapy , Gene Expression/physiology , Humans , Mass Spectrometry , Mental Disorders/physiopathology , Mental Disorders/therapy , Oligonucleotide Array Sequence Analysis , Polymerase Chain Reaction , RNA, Messenger/genetics , Sequence Analysis, DNA , Spectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationABSTRACT
The main active form of atrial natriuretic peptide (ANP) in the circulation is alpha ANP(1-28) [gamma-ANP(99-126)], although some active and inactive metabolites are also though to be present. Some immunoassays for alpha ANP reportedly detect inactive metabolites, as determined by their comparison with results by receptor assay, which would lead to misleading results for alpha ANP(1-28) measurements. Here we compare a radioreceptor assay (RRA) and a radioimmunoassay (RIA) for alpha ANP and show that their results correlate very well with regard to specificity and that the RIA appears to detect only the active circulating form of alpha ANP(1-28).
Subject(s)
Atrial Natriuretic Factor/analysis , Peptide Fragments/analysis , Adrenal Glands/metabolism , Animals , Cross Reactions , Humans , Radioimmunoassay , Radioligand Assay , Receptors, Cell Surface/metabolismSubject(s)
Asialoglycoproteins , alpha-Fetoproteins/metabolism , Animals , Cell Line , Cricetinae , Fetuins , Kidney , Kinetics , Liver , RatsABSTRACT
A foetal-calf-serum component has been isolated that stimulates cells in culture to adopt typical fibroblast morphology. This "spreading" factor is a glycoprotein with a mol. wt. of approx. 62000 and contains 12% carbohydrate; it has an isoelectric point of 4.8. The method described allows purification to homogeneity as judged by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and analytical isoelectric focusing. Under cell-culture conditions the factor is active at 18 micrograms/ml.
