SYPRO Orange is similar to SYPRO Red although it is somewhat brighter and gives slightly higher background fluorescence. For those using a laser-excited gel scanner, an argon-ion laser-based instrument is recommended for SYPRO Orange. The dye is efficiently excited by UV or broad band illumination. It is not suitable for staining proteins on blots or in IEF gels and shows reduced sensitivity when staining proteins on 2-D gels. SYPRO Orange is:
Highly sensitive. The stain can detect 1 to 2 ng of protein per minigel band, making it more sensitive than Coomassie Brilliant Blue or silver staining.
Rapid. Staining is complete in less than one hour
Simple. After electrophoresis, the gel is stained, rinsed and photographed; no fixation or destaining steps are required
Compatible with standard laboratory equipment. Stained proteins can be visualized with a standard 300 nm UV transilluminator or laser scanner.
Cost-effective. Staining with SYPRO Orange is less expensive than silver staining and requires less time.
Low protein-to-protein variability. The dye interacts with the SDS coat around the protein, giving more consistent staining between different types of proteins compared to Coomassie or silver staining.
Selective for proteins. SYPRO Orange detects proteins as small as 6.5 kDa and does not stain nucleic acids or lipopolysaccharides. It does stain glycosylated proteins.
Broad linear range of detection. The fluorescence intensity of the stained bands is linear with protein quantity over three orders of magnitude (a much broader range than either Coomassie or silver staining).
SYPRO orange protein gel stain has been used in the thermal denaturation assay and thermal shift assay of protein(s) of interest. It has also been used in differential scanning fluorimetry.
The SYPRO stock solutions should be stored desiccated and protected from light at room temperature, 4 °C or 20 °C. When stored properly, these stock solutions are stable for six months to one year. The staining reagent diluted in buffer or acetic acid solution can be stored in very clean and detergent-free glass or plastic bottles, protected from light at 4 °C for at least three months.
SYPRO is a registered trademark of Life Technologies
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