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A simple method to remove contaminating salt from IPG strips prior to IEF.
Electrophoresis. 2007 Nov; 28(21):3988-91.E

Abstract

In this study, Saccharomyces cerevisiae yeast lysate is used to demonstrate how a simple wash procedure can improve IEF of IPG strips passively rehydrated in the presence of NaCl. By performing three 10 min washes after IPG strip rehydration and before IEF, corresponding second-dimensional gels from strips containing NaCl look similar to control strips while the second-dimensional gels of unwashed strips contains streaks and spaces devoid of protein. Up to 500 mM NaCl was added to the yeast lysate and successfully focused following this wash regime. Protein loss due to the washes was determined to be minimal by comparing replicates of washed and unwashed strips and analyzing the densities of their corresponding second-dimensional gel spots. In the event of unknown salt contamination, indicated by low voltage during focusing, it is possible to stop focusing, wash the strips, and then continue focusing with acceptable second dimension results.

Authors+Show Affiliations

Heppelmann CJ
Mayo Proteomics Research Center, Mayo Clinic College of Medicine, Rochester, MN 55902, USA.
Benson LM
No affiliation info available
Bergen HR
No affiliation info available

MeSH

BuffersChemical FractionationElectrophoresis, Gel, Two-DimensionalHydrogen-Ion ConcentrationIsoelectric FocusingProteomicsSaccharomyces cerevisiaeSaccharomyces cerevisiae ProteinsSodium Chloride

Pub Type(s)

Journal Article

Language

eng

PubMed ID

17922508

Citation

Heppelmann, Carrie J., et al. "A Simple Method to Remove Contaminating Salt From IPG Strips Prior to IEF." Electrophoresis, vol. 28, no. 21, 2007, pp. 3988-91.
Heppelmann CJ, Benson LM, Bergen HR. A simple method to remove contaminating salt from IPG strips prior to IEF. Electrophoresis. 2007;28(21):3988-91.
Heppelmann, C. J., Benson, L. M., & Bergen, H. R. (2007). A simple method to remove contaminating salt from IPG strips prior to IEF. Electrophoresis, 28(21), 3988-91.
Heppelmann CJ, Benson LM, Bergen HR. A Simple Method to Remove Contaminating Salt From IPG Strips Prior to IEF. Electrophoresis. 2007;28(21):3988-91. PubMed PMID: 17922508.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A simple method to remove contaminating salt from IPG strips prior to IEF. AU - Heppelmann,Carrie J, AU - Benson,Linda M, AU - Bergen,H Robert,3rd PY - 2007/10/9/pubmed PY - 2008/2/27/medline PY - 2007/10/9/entrez SP - 3988 EP - 91 JF - Electrophoresis JO - Electrophoresis VL - 28 IS - 21 N2 - In this study, Saccharomyces cerevisiae yeast lysate is used to demonstrate how a simple wash procedure can improve IEF of IPG strips passively rehydrated in the presence of NaCl. By performing three 10 min washes after IPG strip rehydration and before IEF, corresponding second-dimensional gels from strips containing NaCl look similar to control strips while the second-dimensional gels of unwashed strips contains streaks and spaces devoid of protein. Up to 500 mM NaCl was added to the yeast lysate and successfully focused following this wash regime. Protein loss due to the washes was determined to be minimal by comparing replicates of washed and unwashed strips and analyzing the densities of their corresponding second-dimensional gel spots. In the event of unknown salt contamination, indicated by low voltage during focusing, it is possible to stop focusing, wash the strips, and then continue focusing with acceptable second dimension results. SN - 0173-0835 UR - https://www.unboundmedicine.com/medline/citation/17922508/A_simple_method_to_remove_contaminating_salt_from_IPG_strips_prior_to_IEF_ L2 - https://doi.org/10.1002/elps.200700252 DB - PRIME DP - Unbound Medicine ER -