Literature DB >> 22585508

Long-term, buffer-less, wet gel storage in non-sealed polyethylene bags.

Biji T Kurien1, R Hal Scofield.   

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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Year:  2012        PMID: 22585508      PMCID: PMC7310580          DOI: 10.1007/978-1-61779-821-4_37

Source DB:  PubMed          Journal:  Methods Mol Biol        ISSN: 1064-3745


  8 in total

1.  High resolution two-dimensional electrophoresis of proteins.

Authors:  P H O'Farrell
Journal:  J Biol Chem       Date:  1975-05-25       Impact factor: 5.157

2.  Long-term wet storage of sodium dodecyl sulfate-polyacrylamide gels in polyethylene bags.

Authors:  B T Kurien; R H Scofield
Journal:  Anal Biochem       Date:  1998-07-15       Impact factor: 3.365

3.  Multiple immunoblots after non-electrophoretic bidirectional transfer of a single SDS-PAGE gel with multiple antigens.

Authors:  B T Kurien; R H Scofield
Journal:  J Immunol Methods       Date:  1997-06-23       Impact factor: 2.303

4.  Cleavage of structural proteins during the assembly of the head of bacteriophage T4.

Authors:  U K Laemmli
Journal:  Nature       Date:  1970-08-15       Impact factor: 49.962

5.  Electrophoretic transfer of proteins from fixed and stained gels.

Authors:  D S Phelps
Journal:  Anal Biochem       Date:  1984-09       Impact factor: 3.365

6.  Heat mediated quick Coomassie blue protein staining and destaining of SDS-PAGE gels.

Authors:  B T Kurien; R H Scofield
Journal:  Indian J Biochem Biophys       Date:  1998-12       Impact factor: 1.918

7.  Association of neutropenia in systemic lupus erythematosus (SLE) with anti-Ro and binding of an immunologically cross-reactive neutrophil membrane antigen.

Authors:  B T Kurien; J Newland; C Paczkowski; K L Moore; R H Scofield
Journal:  Clin Exp Immunol       Date:  2000-04       Impact factor: 4.330

8.  Affinity immunoblotting for analysis of antibody clonotype distribution in a lupus patient developing anti-Ro 60 over time.

Authors:  Biji T Kurien; R Hal Scofield
Journal:  Methods Mol Biol       Date:  2009
  8 in total

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