Abstract
We describe a method for the detection of total protein patterns on polyvinylidene difluoride (PVDF) membranes using the fluorogenic dye 2-methoxy-2,4-diphenyl-3(2H)-furanone (MDPF) (1). This method is based on the fluorescent properties of this dye (2). As can be seen in Fig. 1, MDPF (A) and the hydrolysis product (C) are nonfluorescent; only the adduct B formed with the proteins is fluorescent. This makes unnecessary the destaining of the PVDF membrane after protein labeling. The whole process of staining with MDPF is completed in about 20 min. Wet membranes are translucent, allowing the visualization of MDPF labeled protein bands by transillumination with UV light (see Fig. 2A). Electrophoretic bands containing less than 10 ng of protein transferred to PVDF membranes can be detected after the reaction with MDPF.
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References
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Acknowledgment
This work was supported in part by grant BFU2005-3883 (Ministerio de Educación y Ciencia).
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© 2009 Springer Science+Business Media, LLC
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Alba, F.J., Daban, JR. (2009). MDPF Staining of Proteins on Western Blots. In: Walker, J.M. (eds) The Protein Protocols Handbook. Springer Protocols Handbooks. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59745-198-7_71
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DOI: https://doi.org/10.1007/978-1-59745-198-7_71
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60327-474-6
Online ISBN: 978-1-59745-198-7
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