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Rapid detection of proteins by enzyme‐linked immunofiltration assay after transfer onto nitrocellulose membranes

 

作者: Jean‐Michel Pinon,   Dominique Puygauthier‐Toubas,   Hervé Lepan,   Cathy Marx,   Annie Bonhomme,   Joselyne Boulant,   RÉGine Geers,   Hervé Dupont,  

 

期刊: ELECTROPHORESIS  (WILEY Available online 1990)
卷期: Volume 11, issue 1  

页码: 41-45

 

ISSN:0173-0835

 

年代: 1990

 

DOI:10.1002/elps.1150110110

 

出版商: Wiley Subscription Services, Inc., A Wiley Company

 

数据来源: WILEY

 

摘要:

AbstractEnzyme‐linked immunofiltration assay (ELIFA) for labeling transferred proteins is an interesting and powerful technique for the rapid specific detection (15 min) of proteins immobilized on nitrocellulose or nylon membranes (0.20 and 0.45 μm). ELIFA does not require fastidious handling of the membranes. Saturation, specific labeling and washing procedures are achieved by filtration, controlled by a monitoring unit which regulates the flow rate and ensures excellent specificity, repetition and reproducibility. The recycling by closed circuit or by repetetive inversion of the flow direction offers the advantage of reducing the volumes of expensive reagents while simultaneously increasing the sensitivity of the technique. The detection limit is at least as low as 1–5 ng using directly or indirectly enzymatically labelled probes. ELIFA may be extended to the identification of glycoproteins using specific ligands such as lectins or to the immunocapture of an antigen using specific antibodies immobilized on an activated membrane. ELIFA complements fast separation, bye.g., isoelectric focusing, polyacrylamide gel electrophoresis, or sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and accelerated electrotransfer to membranes with rapid detection reducing the total time for separation transfer and detection to less th

 

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