hydrogen 9-(2,4-disulphonatophenyl)-3,6-bis(ethylamino)-2,7-dimethylxanthylium, monosodium salt

Sulforhodamine G is a fluorescent stain with broad dynamic ranges. Sulforhodamine G can be used for the research of protein stains[1].

Research Areas >> Others

Signaling Pathways >> Others >> Others

Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs). Labeling of fluorescent internal protein: A. Prepared of protein samples: 1. Protein samples to be analyzed are spiked with 0.1% of the total protein load of ALIS647 (ALIS internal standard) prior. 2. Protein sample is separated with 2-DE in the dark. B. Purify Sulforhodamine G: 1. Sulforhodamine G (60% purity, 10 mg) dissolved in 100 mL of 1% v/v acetic acid to purify by RP chromatography. 2. Collect the pool with an absorbance maximum at 528 nm, lyophilized to dryness and stored as a dry powder at 47℃. C. Stainning: 1. staining was performed in polypropylene staining dishes wrapped in aluminum foil to prevent photobleaching of the stains. 2. Sulforhodamine G staining is performed overnight in 35% methanol with a four-fold molar excess of dye: protein based on an average protein molecular weight of 50 kDa. 3. Following 4×15 min washes in 35% methanol and 2×15 min equilibrations in water. 4. Total protein and ALIS are visualized with a laser scanner using different channels. 5. Protein spots visualized using total protein stain (λex=532 nm) and ALIS (λex=633 nm) are quantified separately using 2-DE software, and statistical analysis.

[1]. Asa M Wheelock, et al. Use of a fluorescent internal protein standard to achieve quantitative two-dimensional gel electrophoresis. Proteomics. 2006 Mar;6(5):1385-98.