SYPRO Ruby stained Protein gel

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 RubySYPRO OrangeSYPRO RedSYPRO TangerineCoomassie Fluor OrangeSYPRO Ruby Blot StainNo-Stain Protein Labeling Reagent
Min. protein detected0.25- 1 ng4–8 ng4–8 ng1-4 ng8 ng2-8 ng20 ng
Stain time90 min microwave
18 hr standard
~1 hour~1 hour~1 hour~1 hour~1 hour10 min
Ex/EmEx: 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 applications1D and 2D gels (SDS and non-denaturing), IEF, Mass specSDS gel stainingSDS gel stainingSDS gel staining, Western blotting, zymography, mass spec, electroelutionSDS gel stainingMembrane staining, mass spec, microsequencingGel or membrane staining, western blotting
HighlightsStains most classes of proteins, including glycoproteins, phosphoproteins, lipoproteins, calcium binding proteins, fibrillar proteins, and other proteins that are difficult to stainStain can be dissolved in the cathode (top) running buffer to stain proteins as the gel runsStain can be dissolved in the cathode (top) running buffer to stain proteins as the gel runsStaining is performed in buffered salt solutions (no acids or organic solvents required)Gels do not need to be wash before stainingStain does not interfere with subsequent detection techniquesTotal 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
  

  

FormatReady to useSupplied as stock solutionSupplied as stock solutionSupplied as stock solutionReady to useReady to useSupplied as stock solution
Catalog numberS12000S6651S6653S12010C33250S11791A44449

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 RubySYPRO OrangeSYPRO RedSYPRO TangerineCoomassie Fluor OrangeSYPRO Ruby Blot StainNo-Stain Protein Labeling Reagent
Mode of actionNon-covalently binds to basic amino acids and the polypeptide backboneNon-covalently interacts with the SDS coat around proteins in the gelNon-covalently interacts with the SDS coat around proteins in the gelNon-covalently interacts with the SDS coat around proteins in the gelNon-covalently interacts with the SDS coat around proteins in the gelNon-covalently interacts with proteins on the membraneForms 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

 ReagentOvernight protocolMicrowave protocol
Fix50% methanol, 7% acetic acid100 ml, 30 min100 ml, 15 min
100 ml, 30 min100 ml, 15 min
StainSYPRO Ruby gel stain75 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)
Wash10% methanol, 7% acetic acid100 ml, 30 min100 ml, 30 min
Total time ~18 hours90 minutes
Hands on time 10 minutes15 minutes

Staining protocol for SYPRO Orange and SYPRO Red

 ReagentStep
PrepareDilute stock reagent 1:5000 in 7.5% (v/v) acetic acid 
StainPlace gel in staining solution50 mL (mini gel),
10 to 60 min
RinseRinse 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 for gel documentation of fluorescent protein gel and membrane stains

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 for visualization of fluorescent protein gel and membrane stains

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

Compare available fluorescent protein gel and membrane stains

 SYPRO RubySYPRO OrangeSYPRO RedSYPRO TangerineCoomassie Fluor OrangeSYPRO Ruby Blot StainNo-Stain Protein Labeling Reagent
Min. protein detected0.25- 1 ng4–8 ng4–8 ng1-4 ng8 ng2-8 ng20 ng
Stain time90 min microwave
18 hr standard
~1 hour~1 hour~1 hour~1 hour~1 hour10 min
Ex/EmEx: 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 applications1D and 2D gels (SDS and non-denaturing), IEF, Mass specSDS gel stainingSDS gel stainingSDS gel staining, Western blotting, zymography, mass spec, electroelutionSDS gel stainingMembrane staining, mass spec, microsequencingGel or membrane staining, western blotting
HighlightsStains most classes of proteins, including glycoproteins, phosphoproteins, lipoproteins, calcium binding proteins, fibrillar proteins, and other proteins that are difficult to stainStain can be dissolved in the cathode (top) running buffer to stain proteins as the gel runsStain can be dissolved in the cathode (top) running buffer to stain proteins as the gel runsStaining is performed in buffered salt solutions (no acids or organic solvents required)Gels do not need to be wash before stainingStain does not interfere with subsequent detection techniquesTotal 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
  

  

FormatReady to useSupplied as stock solutionSupplied as stock solutionSupplied as stock solutionReady to useReady to useSupplied as stock solution
Catalog numberS12000S6651S6653S12010C33250S11791A44449

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 RubySYPRO OrangeSYPRO RedSYPRO TangerineCoomassie Fluor OrangeSYPRO Ruby Blot StainNo-Stain Protein Labeling Reagent
Mode of actionNon-covalently binds to basic amino acids and the polypeptide backboneNon-covalently interacts with the SDS coat around proteins in the gelNon-covalently interacts with the SDS coat around proteins in the gelNon-covalently interacts with the SDS coat around proteins in the gelNon-covalently interacts with the SDS coat around proteins in the gelNon-covalently interacts with proteins on the membraneForms 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

 ReagentOvernight protocolMicrowave protocol
Fix50% methanol, 7% acetic acid100 ml, 30 min100 ml, 15 min
100 ml, 30 min100 ml, 15 min
StainSYPRO Ruby gel stain75 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)
Wash10% methanol, 7% acetic acid100 ml, 30 min100 ml, 30 min
Total time ~18 hours90 minutes
Hands on time 10 minutes15 minutes

Staining protocol for SYPRO Orange and SYPRO Red

 ReagentStep
PrepareDilute stock reagent 1:5000 in 7.5% (v/v) acetic acid 
StainPlace gel in staining solution50 mL (mini gel),
10 to 60 min
RinseRinse 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 for gel documentation of fluorescent protein gel and membrane stains

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 for visualization of fluorescent protein gel and membrane stains

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

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