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Invitrogen™ SYPRO™ Protein Gel Stains
Description
SYPRO protein gel stains are sensitive, easy-to-use fluorescent stains for total proteins separated by polyacrylamide gel electrophoresis (PAGE). Stained proteins can be viewed with a standard UV or blue-light transilluminator, or imaging equipment containing the appropriate filters or laser. SYPRO protein gel stains offer many advantages over Coomassie Blue and silver staining, including a fast, one-step staining protocol requiring no destaining, a linear dynamic range over three orders of magnitude, and very little protein-to-protein variation in staining.
SYPRO Ruby Protein Gel Stain features include:
- Convenient—ready-to-use formulation
- High sensitivity—detection to 0.25 to 1 ng
- Effective staining for 1D and 2D gels
- Can be multiplexed with other gel stains such as Pro-Q Diamond phosphoprotein gel stain or Pro-Q Emerald 300 glycoprotein gel stain
- Compatible with subsequent analysis of proteins by Edman-based sequencing or mass spectrometry
SYPRO Orange and SYPRO Red protein gel stain features include:
- Sensitive—detection to 4 to 8 ng
- Fast—rapid staining time of 10–60 min
- Best for 1D gels
- SYPRO Orange: Ex: 300, 470 nm, Em: 570 nm
- SYPRO Red: Ex: 300, 550 nm, Em: 630 nm
SYPRO Tangerine Protein Gel Stain features include:
- Sensitive—detection to 4 to 8 ng
- Fast—rapid staining time of 10 to 60 minutes
- Best for 1D gels
- Does not require fixation with acetic acid
- Compatible with subsequent gel staining, western blotting, zymography, electroelution or mass spectrometry
Order Info
Shipping Condition: Room Temperature
Specifications
Specifications
| Content And Storage | Supplied as a 5000X concentrate in DMSO. Store at room temperature and protect from light. |
| Detection Location | In-Gel Detection |
| Color | Orange |
| Concentration | 5000X in DMSO |
| Detection Method | Fluorescence |
| Label or Dye | SYPRO Orange |
| Quantity | 500 μL |
| Shipping Condition | Room Temperature |
| Target Molecule | Protein |
| Excitation/Emission | 300, 470/570 nm |
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Frequently Asked Questions (FAQs)
No. Dye concentrations higher than 1X do not give better detection. Instead, background fluorescence increases and, as the dye concentration increases, the dye becomes self-quenching and the signal actually decreases.
Gels stained with SYPRO dyes can be dried between sheets of cellophane, although there is sometimes a slight decrease in sensitivity. If the gels are dried onto paper, the light will scatter and the sensitivity will decrease. Other plastics are not recommended, as the plastic typically used is not transparent to UV light.
Yes, you may use SYPRO Orange Protein Gel Stain with Invitrogen gels but the protocol would have to be modified. The stain is mainly to be used with 0.05% SDS running buffer while all our buffers contain 0.1% SDS. This results in high background as the SDS binds tightly to the stain. The following protocol is recommended for best results with Invitrogen gels:
Wash solution: 7.5% (v/v) Acetic acid
Staining solution: 1:5000 SYPRO Orange in Wash solution
After the gel run is over, wash the gel in 100 ml of Wash solution for 10 minutes.
Place gel in Staining solution for up to 24 h in the dark.
Remove the gel from the Staining solution and rinse briefly in 100 ml Wash solution.
The gel is then ready to be photographed.
No. Loading solutions contain so much SDS that SYPRO Ruby, SYPRO Orange and SYPRO Red dyes simply localize in the free SDS and bind very little of the proteins. Proteins can be covalently pre-labeled with ATTO-TAG CBQCA (Cat. No. A2333), DDAO succinimidyl ester (Cat. No. C34553) or TAMRA-succinimidyl ester (Cat. No. C2211) dyes, or the TC-FLAsH Expression Analysis Detection Kits (Cat. No. A10067 for orange fluorescence, Cat. No. A10068 for red fluorescence) prior to electrophoresis without affecting protein migration through the gel.
SYPRO Orange or SYPRO Red Protein Gel Stain can be diluted 5000-fold into the cathode (upper) buffer tank to stain proteins during electrophoresis without affecting migration. The problem with doing this is that there is considerable background fluorescence in the gels from the dye interacting with SDS. This can be reduced after electrophoresis by destaining in 7.5% acetic acid for 15-60 minutes. This method also results in poorer protein sensitivity than the standard post-staining method, requires the same amount of time before the gel can be imaged, and contaminates the electrophoresis apparatus.