A Fast and Sensitive Method for Detection of Phospholipid-Binding Proteins on Nitrocellulose Membranes

We describe a sensitive new method for specific detection and quantitation of phospholipid-binding proteins using two purified lipocortins as model proteins. The method consists of blotting proteins to nitrocellulose membranes followed by incubation with radiolabeled phospholipids and autoradiograph...

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Bibliographic Details
Published inAnalytical biochemistry Vol. 209; no. 2; pp. 251 - 257
Main Authors Weber, G., Schwamberger, G., Ferber, E.
Format Journal Article
LanguageEnglish
Published San Diego, CA Elsevier Inc 01.03.1993
Elsevier
Subjects
Animals
Annexins - analysis
Biological and medical sciences
Biotechnology
Collodion
Electrophoresis, Polyacrylamide Gel
Fundamental and applied biological sciences. Psychology
Lung - chemistry
Membrane Proteins - analysis
Membranes, Artificial
Methods. Procedures. Technologies
Mice
Mice, Inbred C57BL
Others
Phospholipids - analysis
Protein Binding
Sensitivity and Specificity
Sodium Dodecyl Sulfate
Swine
Various methods and equipments
Radiolabelling
Gel electrophoresis
Blotting assay
Cellulose nitrate
Lipocortin
Phospholipid
Detection
Quantitative analysis
Calcium Cations
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Summary:We describe a sensitive new method for specific detection and quantitation of phospholipid-binding proteins using two purified lipocortins as model proteins. The method consists of blotting proteins to nitrocellulose membranes followed by incubation with radiolabeled phospholipids and autoradiographic detection of bound phospholipids. It allows specific detection of phospholipid binding proteins to a threshold of about 100 ng/spot and their quantitation up to several micrograms as well as rapid analysis of specific phospholipid binding properties of individual proteins. If performed as a dot-blotting technique, the method permits the simultaneous analysis of a large number of samples and the establishment of Ca2+ and phospholipid binding curves. If blotting is preceded by electrophoresis, the method also allows specific detection and quantitative estimation of phospholipid binding proteins in crude biological preparations and may thus be useful for monitoring and analysis of these proteins in various biological samples.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
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ISSN:0003-2697
1096-0309
DOI:10.1006/abio.1993.1116