SYPRO Ruby staining

Solutions:

SYPRO Ruby protein gel stain, ’Bio-Rad’,
No 170-3125, 170-3126, and 170-3138

Fix solution: 10 MeOH, 7 acetic acid

 

Protocol:

  1. Incubate the gel for 30 min in Fix solution:

 250 ml for a 2-D gel (18.5 20 cm)

  1. Cover the gel with the stain: : 350-400 ml for a gel.

Incubate the gel with gentle agitation on an orbital shaker for at least 3 hr or overnight.

  1. Wash the gel in Fix solution for 30-60 min to reduce background fluorescence.

  2. Wash the gel in water.

Stained proteins can be visualized using a 300 nm UV or a blue-light transilluminator. The excitation maxima are at: 280 nm and 450 nm, and emission maximum at 610 nm.

Important! Use polypropylene or polyvinyl chloride staining containers instead of glass dishes!

2003 06 23 – Julius