Bio-Rad Fluorescent Protein Stains - Manual (Page 6)

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Section 2
Instructions
2.1 General Considerations
Best results are obtained using clean technique. Any dust or dirt transferred to the
surface of the gel may appear in the fluorescence image as smudges or speckles.
Flamingo fluorescent gel stain is exceptionally sensitive. Contaminant proteins
such as keratin will appear in the gel image if care is not taken to minimize such
contamination.
All glassware used should be cleaned with laboratory glassware cleaner and
rinsed with distilled or deionized water. Use dust-free gloves and limit dust
exposure by keeping reagent vessels and gel trays covered as much as possible.
If gels are cast in the laboratory, the glass plates used should be thoroughly
cleaned with lint-free laboratory wipes.
Flamingo fluorescent gel stain may be exposed to typical room light for up to 8 hours
without any observable effect on the fluorescence intensity of the stained protein
bands or spots. However, if gels are ok as is left in stain solution for more extended
periods, photobleaching may occur. This can be prevented by covering the gel
trays with aluminum foil or an opaque lid.
Instructions given are for standard 1 mm thick SDS-PAGE gels. Thicker gels may
benefit from longer fix and stain times and larger volumes of solution. Native PAGE
gels may also be stained with this procedure.
2.2 Prepare Fix Solution
The fix solution consists of 40% (v/v) ethanol and 10% (v/v) acetic acid. Prepare
the solution with distilled or deionized water that has been filtered. The quantity of
fix solution required depends on the number of gels to be stained and the size of
the gel as indicated in the table below.
Gel Size
Volume of Fix Solution
per Gel
Ready Gel (8.6 cm × 6.8 cm) 100 ml
Criterion (13.3 cm × 8.7 cm) 200 ml
PROTEAN II (16 cm × 16 cm) 500 ml
PROTEAN Plus (25.6 cm × 23 cm) 1,000 ml
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