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 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: 510 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
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

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: 510 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
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
Resources

Article: Protein Detection on Gels, Blots and Arrays- Fluorescent Stains

Download Protein Gel Electrophoresis Technical Handbook

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