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Improved method for identification of proteins using two-dimensional electrophoresis with immobilized pH gradient isoelectric focusing.
Electrophoresis. 1991 Sep; 12(9):678-80.E

Abstract

Recent advances in protein sequence analysis now permit the determination of partial N-terminal and internal primary structure from low picomole quantities of protein. The major remaining hurdles to sequence analysis of small amounts of protein are the identification, isolation, and handling of microgram and submicrogram quantities of protein. The technique of two-dimensional electrophoresis using immobilized pH gradient isoelectric focusing circumvents many of these problems. However, poor correlation between the first and second dimension have prevented use of this technique for the identification of some proteins which can only be assayed prior to the denaturing conditions used in the second dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis procedure. An improved method is presented which allows correlation of the native biological activity (first dimension) to a silver stained protein (second dimension) with a high degree of confidence.

Authors+Show Affiliations

Department of Molecular Neuroendocrinology, Max Planck Institute for Experimental Medicine, Göttingen.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

1752252

Citation

Mackin, R B., et al. "Improved Method for Identification of Proteins Using Two-dimensional Electrophoresis With Immobilized pH Gradient Isoelectric Focusing." Electrophoresis, vol. 12, no. 9, 1991, pp. 678-80.
Mackin RB, Raker VA, Spiess J. Improved method for identification of proteins using two-dimensional electrophoresis with immobilized pH gradient isoelectric focusing. Electrophoresis. 1991;12(9):678-80.
Mackin, R. B., Raker, V. A., & Spiess, J. (1991). Improved method for identification of proteins using two-dimensional electrophoresis with immobilized pH gradient isoelectric focusing. Electrophoresis, 12(9), 678-80.
Mackin RB, Raker VA, Spiess J. Improved Method for Identification of Proteins Using Two-dimensional Electrophoresis With Immobilized pH Gradient Isoelectric Focusing. Electrophoresis. 1991;12(9):678-80. PubMed PMID: 1752252.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Improved method for identification of proteins using two-dimensional electrophoresis with immobilized pH gradient isoelectric focusing. AU - Mackin,R B, AU - Raker,V A, AU - Spiess,J, PY - 1991/9/1/pubmed PY - 1991/9/1/medline PY - 1991/9/1/entrez SP - 678 EP - 80 JF - Electrophoresis JO - Electrophoresis VL - 12 IS - 9 N2 - Recent advances in protein sequence analysis now permit the determination of partial N-terminal and internal primary structure from low picomole quantities of protein. The major remaining hurdles to sequence analysis of small amounts of protein are the identification, isolation, and handling of microgram and submicrogram quantities of protein. The technique of two-dimensional electrophoresis using immobilized pH gradient isoelectric focusing circumvents many of these problems. However, poor correlation between the first and second dimension have prevented use of this technique for the identification of some proteins which can only be assayed prior to the denaturing conditions used in the second dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis procedure. An improved method is presented which allows correlation of the native biological activity (first dimension) to a silver stained protein (second dimension) with a high degree of confidence. SN - 0173-0835 UR - https://www.unboundmedicine.com/medline/citation/1752252/Improved_method_for_identification_of_proteins_using_two_dimensional_electrophoresis_with_immobilized_pH_gradient_isoelectric_focusing_ DB - PRIME DP - Unbound Medicine ER -
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