Abstract
Electrophoresed gels are normally fixed, stained, destained, and dried. Drying is normally carried out with commercial gel dryers or by drying between two cellophane sheets held together by two acrylic frames. Here, we report that stained and destained gels (7.5, 10, or 15%, denaturing or native gels; 0.4 or 1.5 mm in thickness) could be stored wet, unsealed, and without any storage buffer for several months at room temperature within flexible polyethylene bags without significant shrinking or protein diffusion. The gel remains hydrated because of the de facto sealing achieved by the polyethylene sheets (PS) adhering airtightly to the gel on either sides. The microsaturated environment generated by the thin film of water molecules trapped between the gel and the PS, along with the nonporous nature of the PS, apparently protects the gel from cracking as well as shrinking significantly. The intensity of stained proteins increased during storage probably from the slight gel shrinkage observed. Wet gel storage is useful (a) when low abundance protein spots from multiple two-dimensional electrophoresis gels have to be excised for in-gel tryptic digestion or electroelution and (b) for wet gel autoradiography. In addition to avoiding dryer contamination and saving drying time, these bags prevent the moist gel from sticking to X-ray film. Such storage could also prove useful for electrophoretic transfer of fixed and stained gels.
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© 2012 Springer Science+Business Media, LLC
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Kurien, B.T., Scofield, R.H. (2012). Long-Term, Buffer-Less, Wet Gel Storage in Non-sealed Polyethylene Bags. In: Kurien, B., Scofield, R. (eds) Protein Electrophoresis. Methods in Molecular Biology, vol 869. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-821-4_37
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DOI: https://doi.org/10.1007/978-1-61779-821-4_37
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Publisher Name: Humana Press, Totowa, NJ
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Online ISBN: 978-1-61779-821-4
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