A cost effective non-commercial ECL-solution for Western blot detections yielding strong signals and low background (Record no. 46736)

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control field 20250625153857.0
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Transcribing agency CICY
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Classification number (OCLC) (R) ; Classification number, CALL (RLIN) (NR) B-12529
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Title A cost effective non-commercial ECL-solution for Western blot detections yielding strong signals and low background
490 0# - SERIES STATEMENT
Volume/sequential designation Journal of Immunological Methods, 318(1-2), p.11-19, 2007
520 3# - SUMMARY, ETC.
Summary, etc. We compared several alternative ECL solutions for Western blot detection of endogenous proteins in whole cell lysates using inexpensive, commercially available reagents. Starting from an existing protocol based on p-coumaric acid (pCA)as enhancer, we found that the ECL solution containing 4-iodophenylboronic acid (4IPBA)generated strong specific signals and low background chemiluminescence. We optimised the luminol, 4IPBA and hydrogenperoxide concentrations of this 4IPBA-ECL solution. The optimised 4IPBA-ECL solution (100 mM Tris/HCl pH 8.8, 1.25 mM luminol, 2 mM 4IPBA, 5.3 mM hydrogenperoxide)shows a greatly increased signal intensity compared to the initial pCA-ECL protocol and to some commercially available ECL solutions. In addition, the optimised 4IPBA-ECL solution also generates much lower background chemiluminescence than other noncommercial ECL solutions using p-coumaric acid or 4-iodophenol as enhancers. The 4IPBA-ECL solution was table when stored but had the lowest background when prepared freshly from stock solutions. Thus, we present an optimised protocol for a wellperforming inexpensive ECL solution which is an alternative to expensive commercial ECL solutions and which achieves a better signal and lower background than the commercial solutions tested.
650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element ENHANCED CHEMILUMINESCENCE
650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element WESTERN BLOT
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Topical term or geographic name entry element 4-IODOPHENYLBORONIC ACID
650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element P-COUMARIC ACID
650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element 4-IODOPHENOL
700 12 - ADDED ENTRY--PERSONAL NAME
Personal name Haan, C.
700 12 - ADDED ENTRY--PERSONAL NAME
Personal name Behrmann, I.
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Uniform Resource Identifier <a href="https://drive.google.com/file/d/1ofQoNnWiZsIk0xw6hyqoSDPrlVy1RKDm/view?usp=drivesdk">https://drive.google.com/file/d/1ofQoNnWiZsIk0xw6hyqoSDPrlVy1RKDm/view?usp=drivesdk</a>
Public note Para ver el documento ingresa a Google con tu cuenta: @cicy.edu.mx
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  Clasificación local     Ref1 CICY CICY Documento préstamo interbibliotecario 25.06.2025   B-12529 25.06.2025 25.06.2025 Documentos solicitados