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 kit Pro-Q Emerald 300 Glycoprotein Gel and Blot Stain kit Pierce Glycoprotein Stain Pro-Q Diamond Phosphoprotein Gel Staining Kit Pro-Q Diamond Phosphoprotein Blot Stain Kit
Protein modification Glycoproteins Glycoproteins Glycoproteins Phosphoproteins Phosphoproteins
Sensitivity 4 ng glycoprotein / band 0.5 ng glycoprotein / band 0.6-160 ng glycoprotein / band 1–16 ng phosphoprotein / band 8-16 ng
Stain/destain ~6 hr ~5 hr 2 hr 40 min 4–5 hr ~75 min
Ex/Em 510/520 nm 280/530 nm Colorimetric 555/580 nm 555/580 nm
Advantages Selective staining of glycoproteins Can be detected with 300nm UV illumination No specialized equipment required for visualization Selective staining of phosphoproteins Linear dynamic range of approximately 15-fold
Mode of action Interactions with periodate-oxidized carbohydrate groups at glycosylation sites Interactions with periodate-oxidized carbohydrate groups at glycosylation sites Periodate-acid-Schiff (PAS) reagent method Detects phosphate groups attached to tyrosine, serine, or threonine residues Detects phosphate groups attached to tyrosine, serine, or threonine residues through non-covalent interactions
Number of protocol steps 6 (8 for blot staining) 6 7 5 6
Components 3 3 3 1 2
Compatible applications In-gel (1D or 2D), in-blot detection (PVDF), mass spectrometry In-gel (1D or 2D), in-blot detection (PVDF), mass spectrometry In-gel (1D or 2D), in-blot detection (nitrocellulose) In-gel (1D or 2D), mass spectrometry In-blot detection (PVDF and nitrocellulose), mass spectrometry
Catalog number P21875 P21857 24562 P33300 P33356

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 Stain Pierce 6xHis Protein Tag Stain Reagent Lumio Green Detection Kit
Target recognition sequence Oligohistidine sequence Oligohistidine sequence tetracysteine Lumio tag (Cys-Cys-Pro-Gly-Cys-Cys)
Detection procedure Post-electrophoresis staining Post-electrophoresis staining Pre-electrophoresis labeling
Length of detection procedure Under 3 hr ~2 hr Immediate–part of regular gel electrophoresis sample preparation
Detection sensitivity Nanogram levels 0.2µg Nanogram levels
Ex/Em 300, 560nm/590nm 280-310nm 500nm/535nm
Compatible downstream applications The gel can be stained with Coomassie, silver, or fluorescent stains for total protein content after InVision staining Coomassie staining, silver staining, fluorescent staining, western blotting, or mass spectrometry analysis Coomassie staining, silver staining, fluorescent staining, western blotting, or mass spectrometry analysis
Catalog number Stain only: LC6030
Kit (includes BenchMark his-tagged standard): LC6033
24570 LC6090

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 at: JBC Online).

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 kit Pro-Q Emerald 300 Glycoprotein Gel and Blot Stain kit Pierce Glycoprotein Stain Pro-Q Diamond Phosphoprotein Gel Staining Kit Pro-Q Diamond Phosphoprotein Blot Stain Kit
Protein modification Glycoproteins Glycoproteins Glycoproteins Phosphoproteins Phosphoproteins
Sensitivity 4 ng glycoprotein / band 0.5 ng glycoprotein / band 0.6-160 ng glycoprotein / band 1–16 ng phosphoprotein / band 8-16 ng
Stain/destain ~6 hr ~5 hr 2 hr 40 min 4–5 hr ~75 min
Ex/Em 510/520 nm 280/530 nm Colorimetric 555/580 nm 555/580 nm
Advantages Selective staining of glycoproteins Can be detected with 300nm UV illumination No specialized equipment required for visualization Selective staining of phosphoproteins Linear dynamic range of approximately 15-fold
Mode of action Interactions with periodate-oxidized carbohydrate groups at glycosylation sites Interactions with periodate-oxidized carbohydrate groups at glycosylation sites Periodate-acid-Schiff (PAS) reagent method Detects phosphate groups attached to tyrosine, serine, or threonine residues Detects phosphate groups attached to tyrosine, serine, or threonine residues through non-covalent interactions
Number of protocol steps 6 (8 for blot staining) 6 7 5 6
Components 3 3 3 1 2
Compatible applications In-gel (1D or 2D), in-blot detection (PVDF), mass spectrometry In-gel (1D or 2D), in-blot detection (PVDF), mass spectrometry In-gel (1D or 2D), in-blot detection (nitrocellulose) In-gel (1D or 2D), mass spectrometry In-blot detection (PVDF and nitrocellulose), mass spectrometry
Catalog number P21875 P21857 24562 P33300 P33356

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 Stain Pierce 6xHis Protein Tag Stain Reagent Lumio Green Detection Kit
Target recognition sequence Oligohistidine sequence Oligohistidine sequence tetracysteine Lumio tag (Cys-Cys-Pro-Gly-Cys-Cys)
Detection procedure Post-electrophoresis staining Post-electrophoresis staining Pre-electrophoresis labeling
Length of detection procedure Under 3 hr ~2 hr Immediate–part of regular gel electrophoresis sample preparation
Detection sensitivity Nanogram levels 0.2µg Nanogram levels
Ex/Em 300, 560nm/590nm 280-310nm 500nm/535nm
Compatible downstream applications The gel can be stained with Coomassie, silver, or fluorescent stains for total protein content after InVision staining Coomassie staining, silver staining, fluorescent staining, western blotting, or mass spectrometry analysis Coomassie staining, silver staining, fluorescent staining, western blotting, or mass spectrometry analysis
Catalog number Stain only: LC6030
Kit (includes BenchMark his-tagged standard): LC6033
24570 LC6090

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 at: JBC Online).

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.

Resources

Article: Detecting Protein Modifications

Download Protein Gel Electrophoresis Technical Handbook

Download Complete, innovative western workflow solutions brochure