Detection of fluorescence dye-labeled proteins in 2-D gels using an Arthur 1442 Multiwavelength Fluoroimager.
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.
Herick
K
K
PerkinElmer Life Sciences/Wallac Oy, Turku, Finland. klaus.herick@perkinelmer.com
Jackson
P
P
Wersch
G
G
Burkovski
A
A
D001426:Bacterial Proteins / Q000032:analysis; D003352:Corynebacterium; D015180:Electrophoresis, Gel, Two-Dimensional / Q000295:instrumentation / Q000379:methods*; D005063:Europium; D005456:Fluorescent Dyes; D012680:Sensitivity and Specificity
D001426:Bacterial Proteins; D005456:Fluorescent Dyes; C436863:SYPRO Rose Plus; D005063:Europium