2D gel stained with Pro-Q Diamond phosphoprotein stain, Pro-Q Emerald glycoprotein stain and SYPRO Ruby total protein stain

We have a suite of compatible methodologies for differential staining for specific proteins modifications (phosphorylation, glycosylation) and functional groups. The direct stains eliminate the need for antibodies or use of radioisotopes. This set of protein stains not only offer the capacity for the simultaneous detection of multiple protein targets in a single sample, but also provide a combination of high sensitivity and simplicity that can streamline protocols in 1D and 2D polyacrylamide gels or on blots. Explore our stains for phosphoproteins, glycoproteins, and fusion tags below.

Compare different protein staining methods

Our protein modification staining kits allow for in-gel and in-blot phosphoprotein and glycoprotein detection. Our phosphoprotein stains are ideal for the identification of kinase targets in signal transduction pathways and for phosphoproteomic studies. Our sensitive glycoprotein staining kits provide linearity and ease of use for selective detection of glycoproteins.

 Pro-Q Emerald 488 Glycoprotein Gel and Blot Stain kitPro-Q Emerald 300 Glycoprotein Gel and Blot Stain kitPierce Glycoprotein StainPro-Q Diamond Phosphoprotein Gel Staining KitPro-Q Diamond Phosphoprotein Blot Stain Kit
Protein modificationGlycoproteinsGlycoproteinsGlycoproteinsPhosphoproteinsPhosphoproteins
Sensitivity4 ng glycoprotein / band0.5 ng glycoprotein / band0.6-160 ng glycoprotein / band1–16 ng phosphoprotein / band8-16 ng
Stain/destain~6 hr~5 hr2 hr 40 min4–5 hr~75 min
Ex/Em510/520 nm280/530 nmColorimetric555/580 nm555/580 nm
AdvantagesSelective staining of glycoproteinsCan be detected with 300nm UV illuminationNo specialized equipment required for visualizationSelective staining of phosphoproteinsLinear dynamic range of approximately 15-fold
Mode of actionInteractions with periodate-oxidized carbohydrate groups at glycosylation sitesInteractions with periodate-oxidized carbohydrate groups at glycosylation sitesPeriodate-acid-Schiff (PAS) reagent methodDetects phosphate groups attached to tyrosine, serine, or threonine residuesDetects phosphate groups attached to tyrosine, serine, or threonine residues through non-covalent interactions
Number of protocol steps6 (8 for blot staining)6756
Components33312
Compatible applicationsIn-gel (1D or 2D), in-blot detection (PVDF), mass spectrometryIn-gel (1D or 2D), in-blot detection (PVDF), mass spectrometryIn-gel (1D or 2D), in-blot detection (nitrocellulose)In-gel (1D or 2D), mass spectrometryIn-blot detection (PVDF and nitrocellulose), mass spectrometry
Catalog numberP21875P2185724562P33300P33356

Two specific in-gel detection and staining techniques—His-tag in-gel stains and Lumio Green Detection Kit enable detection of tagged-fusion proteins directly in a polyacrylamide gel and allow rapid screening or confirmation of protein expression.

In-gel reagents for visualizing tagged-fusion proteins

 InVision His-tag In-gel StainPierce 6xHis Protein Tag Stain ReagentLumio Green Detection Kit
Target recognition sequenceOligohistidine sequenceOligohistidine sequencetetracysteine Lumio tag (Cys-Cys-Pro-Gly-Cys-Cys)
Detection procedurePost-electrophoresis stainingPost-electrophoresis stainingPre-electrophoresis labeling
Length of detection procedureUnder 3 hr~2 hrImmediate–part of regular gel electrophoresis sample preparation
Detection sensitivityNanogram levels0.2µgNanogram levels
Ex/Em300, 560nm/590nm280-310nm500nm/535nm
Compatible downstream applicationsThe gel can be stained with Coomassie, silver, or fluorescent stains for total protein content after InVision stainingCoomassie staining, silver staining, fluorescent staining, western blotting, or mass spectrometry analysisCoomassie staining, silver staining, fluorescent staining, western blotting, or mass spectrometry analysis
Catalog numberStain only: LC6030
Kit (includes BenchMark his-tagged standard): LC6033
24570LC6090

Three technologies—SYPRO Ruby protein gel stain, Pro-Q Diamond phosphoprotein gel stain and the Pro-Q Emerald glycoprotein stains—can be used individually or in series to detect total protein, phosphoproteins, and glycoproteins, respectively, using a single protein sample separated by 1D or 2D gel electrophoresis. Unambiguous spot matching of phosphoproteins and glycoproteins is made simple by direct comparison with the total protein profile stained with SYPRO Ruby stain.

2D gel stained with Pro-Q Diamond phosphoprotein stain, Pro-Q Emerald glycoprotein stain and SYPRO Ruby total protein stain

Multiplex staining: Proteins from either normal liver cells (left) or liver tumor cells (right) were separated using 2D gel electrophoresis and stained with Pro-Q Diamond phosphoprotein gel stain (blue), Pro-Q Emerald 300 glycoprotein stain (green) and then SYPRO Ruby protein gel stain (red). The gel was imaged after each staining and the digital images were overlaid.

Multiplexed proteomics technology for prescreening

Almost all modern proteomics studies involve some type of prefractionation sample analysis. There are often multiple sample fractions to analyze, and it is usually not clear which fractions will have the most interesting proteins. We recommend running all sample fractions on simple 1D gels (10 to 15 fractions per gel), and then staining the 1D gel for phosphoproteins, glycoproteins, and total proteins using the Pro-Q Diamond, Pro-Q Emerald, and SYPRO Ruby protein gel stains, respectively. In this manner, it is possible to quickly determine the presence of phosphorylated and glycosylated proteins in a fraction, in relation to total-protein content (Read more).

Used on its own, each of these state-of-the-art detection technologies brings you the following benefits:

  • Simple, streamlined staining method
  • High sensitivity
  • Broad linear quantitation range
  • Compatibility with many instruments, including gel scanners and mass spectrometers
Analysis of the glycoproteome and phosphoproteome using Pro-Q and SYPRO stains

Multiplexed Proteomics technologies applied to 1 D gels. Differential staining using Pro-Q Diamond, Pro-Q Emerald and SYPRO Ruby was use to visualize and compare total protein, glycoproteins and phosphorptoreins in a normal and tumor sample.

Our protein modification staining kits allow for in-gel and in-blot phosphoprotein and glycoprotein detection. Our phosphoprotein stains are ideal for the identification of kinase targets in signal transduction pathways and for phosphoproteomic studies. Our sensitive glycoprotein staining kits provide linearity and ease of use for selective detection of glycoproteins.

 Pro-Q Emerald 488 Glycoprotein Gel and Blot Stain kitPro-Q Emerald 300 Glycoprotein Gel and Blot Stain kitPierce Glycoprotein StainPro-Q Diamond Phosphoprotein Gel Staining KitPro-Q Diamond Phosphoprotein Blot Stain Kit
Protein modificationGlycoproteinsGlycoproteinsGlycoproteinsPhosphoproteinsPhosphoproteins
Sensitivity4 ng glycoprotein / band0.5 ng glycoprotein / band0.6-160 ng glycoprotein / band1–16 ng phosphoprotein / band8-16 ng
Stain/destain~6 hr~5 hr2 hr 40 min4–5 hr~75 min
Ex/Em510/520 nm280/530 nmColorimetric555/580 nm555/580 nm
AdvantagesSelective staining of glycoproteinsCan be detected with 300nm UV illuminationNo specialized equipment required for visualizationSelective staining of phosphoproteinsLinear dynamic range of approximately 15-fold
Mode of actionInteractions with periodate-oxidized carbohydrate groups at glycosylation sitesInteractions with periodate-oxidized carbohydrate groups at glycosylation sitesPeriodate-acid-Schiff (PAS) reagent methodDetects phosphate groups attached to tyrosine, serine, or threonine residuesDetects phosphate groups attached to tyrosine, serine, or threonine residues through non-covalent interactions
Number of protocol steps6 (8 for blot staining)6756
Components33312
Compatible applicationsIn-gel (1D or 2D), in-blot detection (PVDF), mass spectrometryIn-gel (1D or 2D), in-blot detection (PVDF), mass spectrometryIn-gel (1D or 2D), in-blot detection (nitrocellulose)In-gel (1D or 2D), mass spectrometryIn-blot detection (PVDF and nitrocellulose), mass spectrometry
Catalog numberP21875P2185724562P33300P33356

Two specific in-gel detection and staining techniques—His-tag in-gel stains and Lumio Green Detection Kit enable detection of tagged-fusion proteins directly in a polyacrylamide gel and allow rapid screening or confirmation of protein expression.

In-gel reagents for visualizing tagged-fusion proteins

 InVision His-tag In-gel StainPierce 6xHis Protein Tag Stain ReagentLumio Green Detection Kit
Target recognition sequenceOligohistidine sequenceOligohistidine sequencetetracysteine Lumio tag (Cys-Cys-Pro-Gly-Cys-Cys)
Detection procedurePost-electrophoresis stainingPost-electrophoresis stainingPre-electrophoresis labeling
Length of detection procedureUnder 3 hr~2 hrImmediate–part of regular gel electrophoresis sample preparation
Detection sensitivityNanogram levels0.2µgNanogram levels
Ex/Em300, 560nm/590nm280-310nm500nm/535nm
Compatible downstream applicationsThe gel can be stained with Coomassie, silver, or fluorescent stains for total protein content after InVision stainingCoomassie staining, silver staining, fluorescent staining, western blotting, or mass spectrometry analysisCoomassie staining, silver staining, fluorescent staining, western blotting, or mass spectrometry analysis
Catalog numberStain only: LC6030
Kit (includes BenchMark his-tagged standard): LC6033
24570LC6090

Three technologies—SYPRO Ruby protein gel stain, Pro-Q Diamond phosphoprotein gel stain and the Pro-Q Emerald glycoprotein stains—can be used individually or in series to detect total protein, phosphoproteins, and glycoproteins, respectively, using a single protein sample separated by 1D or 2D gel electrophoresis. Unambiguous spot matching of phosphoproteins and glycoproteins is made simple by direct comparison with the total protein profile stained with SYPRO Ruby stain.

2D gel stained with Pro-Q Diamond phosphoprotein stain, Pro-Q Emerald glycoprotein stain and SYPRO Ruby total protein stain

Multiplex staining: Proteins from either normal liver cells (left) or liver tumor cells (right) were separated using 2D gel electrophoresis and stained with Pro-Q Diamond phosphoprotein gel stain (blue), Pro-Q Emerald 300 glycoprotein stain (green) and then SYPRO Ruby protein gel stain (red). The gel was imaged after each staining and the digital images were overlaid.

Multiplexed proteomics technology for prescreening

Almost all modern proteomics studies involve some type of prefractionation sample analysis. There are often multiple sample fractions to analyze, and it is usually not clear which fractions will have the most interesting proteins. We recommend running all sample fractions on simple 1D gels (10 to 15 fractions per gel), and then staining the 1D gel for phosphoproteins, glycoproteins, and total proteins using the Pro-Q Diamond, Pro-Q Emerald, and SYPRO Ruby protein gel stains, respectively. In this manner, it is possible to quickly determine the presence of phosphorylated and glycosylated proteins in a fraction, in relation to total-protein content (Read more).

Used on its own, each of these state-of-the-art detection technologies brings you the following benefits:

  • Simple, streamlined staining method
  • High sensitivity
  • Broad linear quantitation range
  • Compatibility with many instruments, including gel scanners and mass spectrometers
Analysis of the glycoproteome and phosphoproteome using Pro-Q and SYPRO stains

Multiplexed Proteomics technologies applied to 1 D gels. Differential staining using Pro-Q Diamond, Pro-Q Emerald and SYPRO Ruby was use to visualize and compare total protein, glycoproteins and phosphorptoreins in a normal and tumor sample.