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Detection of Fluorescence Dye-Labeled Proteins in 2-D Gels Using an Arthur™ 1442 Multiwavelength Fluoroimager
oleh: Klaus Herick, Peter Jackson, Gregor Wersch, Andreas Burkovski
Format: | Article |
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Diterbitkan: | Taylor & Francis Group 2001-07-01 |
Deskripsi
Labeling of proteins with SYPRO® Orange, SYPRO Red, and SYPRO Ruby after 2-D polyacrylamide gel electrophoresis (PAGE) using plastic-backed immobilized pH gradient (IPG) strips and precast SDS polyacrylamide gels was tested. Protein spots were detected using an Arthur™ 1442 Multiwavelength Fluoroimager. The labeling methods described allow detection of proteins both after isoelectric focusing (IEF) and PAGE with a sensitivity higher than or comparable to standard silver staining methods. In addition to the post-labeling methods mentioned above, pre-labeling with the cysteine-specific fluorophore monobromobimane before 2-D PAGE is a sensitive, fast, and cost-effective alternative to existing staining protocols.