Fluorescent Protein Gel and Membrane Stains | Thermo Fisher Scientific

Fluorescent stains are rapid, and highly sensitive for detecting total protein in protein electrophoresis gels and membranes. Fluorescent stains are designed for use in 1D and 2D PAGE and offer sensitivities similar to that obtained with silver staining techniques. Our line of SYPRO and fluorescent stains can be used for total protein quantitation and can be viewed using a standard UV or blue-light transilluminator or with imaging instruments equipped with appropriate light sources, such as iBright Imaging Systems.

Compare different protein staining methods

Highlights of fluorescent stains:

Simple staining procedure—no destaining or timed steps required; minimal hands-on time
Quantitative—linear quantitation range over 2-3 orders of magnitude with low protein-to-protein variability
Sensitivity—typically more sensitive than coomassie stains and equivalent to silver stains

Compare available fluorescent protein gel and membrane stains

	SYPRO Ruby	SYPRO Orange	SYPRO Red	SYPRO Tangerine	Coomassie Fluor Orange	SYPRO Ruby Blot Stain	No-Stain Protein Labeling Reagent
Min. protein detected	0.25- 1 ng	4–8 ng	4–8 ng	1-4 ng	8 ng	2-8 ng	20 ng
Stain time	90 min microwave 18 hr standard	~1 hour	~1 hour	~1 hour	~1 hour	~1 hour	10 min
Ex/Em	Ex: 280, 450nm Em: 610 nm	Ex: 300, 470nm Em: 570 nm	Ex: 300, 550nm Em: 630 nm	Ex: 300, 490nm Em: 640 nm	Ex: 300, 470nm Em: 570 nm	Ex: 280, 450nm Em: 618nm	Ex: 488nm Em: 590nm
Compatible applications	1D and 2D gels (SDS and non-denaturing), IEF, Mass spec	SDS gel staining	SDS gel staining	SDS gel staining, Western blotting, zymography, mass spec, electroelution	SDS gel staining	Membrane staining, mass spec, microsequencing	Gel or membrane staining, western blotting
Highlights	Stains most classes of proteins, including glycoproteins, phosphoproteins, lipoproteins, calcium binding proteins, fibrillar proteins, and other proteins that are difficult to stain	Stain can be dissolved in the cathode (top) running buffer to stain proteins as the gel runs	Stain can be dissolved in the cathode (top) running buffer to stain proteins as the gel runs	Staining is performed in buffered salt solutions (no acids or organic solvents required)	Gels do not need to be wash before staining	Stain does not interfere with subsequent detection techniques	Total protein normalization after western blotting.
No fixative required		✓	✓	✓	✓	Fixative required only for PVDF membranes	✓
No de-staining required		✓	✓	✓	✓	✓	✓
Multiplexing with other stains	✓ *not recommended with colorimetric stains			✓			✓
Format	Ready to use	Supplied as stock solution	Supplied as stock solution	Supplied as stock solution	Ready to use	Ready to use	Supplied as stock solution
Catalog number	S12000	S6651	S6653	S12010	C33250	S11791	A44449

Total protein detection

SYPRO stains provide total protein detection by binding non-covalently to proteins in the gel or membrane. Since proteins are not covalently modified by the dyes, the stains are compatible with downstream applications such as mass spectrometry, and sequencing. No-Stain Protein Labeling Reagent covalently modifies proteins in the gel or on the membrane to provide total protein detection.

	SYPRO Ruby	SYPRO Orange	SYPRO Red	SYPRO Tangerine	Coomassie Fluor Orange	SYPRO Ruby Blot Stain	No-Stain Protein Labeling Reagent
Mode of action	Non-covalently binds to basic amino acids and the polypeptide backbone	Non-covalently interacts with the SDS coat around proteins in the gel	Non-covalently interacts with the SDS coat around proteins in the gel	Non-covalently interacts with the SDS coat around proteins in the gel	Non-covalently interacts with the SDS coat around proteins in the gel	Non-covalently interacts with proteins on the membrane	Forms covalent bonds with lysine amino acid side chains with proteins in the gel or membrane

Fluorescent detection

All the SYPRO stains have dual-excitation maxima, the dye exhibits luminescence upon excitation with either UV or visible light. This property makes it possible to visualize the luminescence with many types of instruments, including UV epi-illumination sources, UV or blue-light transilluminators, laser-scanning instruments, and CCD imaging instruments, including those with excitation light at 450 nm, 473 nm, 488 nm or 532 nm. SYPRO stains have exceptional photo stability, allowing long exposure times for maximum sensitivity.

Excitation and emission spectra for SYPRO Ruby Protein Stain

Excitation (dashed line) and emission (solid line) spectra for SYPRO Ruby protein gel stain.

Simple staining protocols

The SYPRO stains follow simple staining procedures which can be completed in less than an hour for a majority of the experiments. SYPRO Orange, Red, and Tangerine require no separate fixation or de staining steps and there is no fear of overstaining the gel.

Staining protocol for SYPRO Ruby Gel Stain

	Reagent	Overnight protocol	Microwave protocol
Fix	50% methanol, 7% acetic acid	100 ml, 30 min	100 ml, 15 min
Fix	50% methanol, 7% acetic acid	100 ml, 30 min	100 ml, 15 min
Stain	SYPRO Ruby gel stain	75 ml, overnight (room temperature)	75 ml, microwave 30 seconds, agitate 30 seconds, microwave 30 seconds, agitate 5 minutes, microwave 30 seconds, agitate 23 minutes (30 minutes total) (80-85 °C)
Wash	10% methanol, 7% acetic acid	100 ml, 30 min	100 ml, 30 min
Total time		~18 hours	90 minutes
Hands on time		10 minutes	15 minutes

Staining protocol for SYPRO Orange and SYPRO Red

	Reagent	Step
Prepare	Dilute stock reagent 1:5000 in 7.5% (v/v) acetic acid
Stain	Place gel in staining solution	50 mL (mini gel), 10 to 60 min
Rinse	Rinse gel in 7.5% acetic acid	<1 min

Visualization, capture and analysis of fluorescent protein gel and membrane stains

All of the SYPRO stains have dual-excitation maxima—the dyes exhibit luminescence upon excitation with either UV or visible light. This property makes it possible to visualize the light emission with many types of instruments, including UV epi-illumination sources, UV or blue light transilluminators, laser-scanning instruments, and CCD imaging instruments, including those that provide excitation light at 450 nm, 473 nm, 488 nm or 532 nm.

iBright Imaging Systems

Capture and analyze publication quality images from fluorescently stained gels and membranes with iBright Imaging Systems. These high-performance instruments enhance the imaging experience through powerful hardware, advanced automated technologies, and an interface that is easy to use for researchers of all experience levels.

Explore our iBright gel documentation systems

Blue light transilluminators

Blue LED transilluminators provide quick, safe way to visualize fluorescently stained gels and membranes. Explore our options for blue light transilluminators.

Explore blue light transilluminators

Overview