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Sequential Double Fluorescent Detections of Total Proteins and Phosphoproteins in SDS-PAGE

Sequential Double Fluorescent Detections of Total Proteins and Phosphoproteins in SDS-PAGE

Sun-Young Hwang, Xu Wang, Wei-Tao Cong, Li-Tai Jin, Jung-Kap Choi

Electrophoresis. 2014 Apr;35(8):1089-98.

PMID: 24488794

Abstract:

A fluorescent staining technique, using selective chelation with fluorophore and metal ion to the phosphate groups of phosphoproteins in SDS-PAGE is described. As a fluorescent dye and a metal ion, Fura 2 pentapotassium salt and Al(3+) were employed, respectively. The staining method, Fura 2 stain, has sensitivities of 16-32 ng of α-casein and β-casein, 62 ng of ovalbumin, phosvitin, and κ-casein using an ultraviolet transilluminator. Furthermore, Fura 2 stain is able to carry out continuative double detection of total proteins and phosphoproteins on the same gel within 3.5 h. Consequently, selective phosphoprotein and total protein detections could be obtained without other poststaining. Considering the low cost, simplicity, and speed, Fura 2 staining may provide great practicalities in routine phosphoproteomics research.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP113694647	Fura 2 pentapotassium salt		113694-64-7	Price

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