Additional Protein Staining Reagents

• Protein Stains Selection Chart
• GlycoProfile™ III Fluorescent Glycoprotein Detection Kit
• Alcian Blue 8GX
• Amido Black Staining Solution, 2X concentrate
• Eosin Y
• Fast Green FCF
• Fluorescamine®
• Glycoprotein Detection Kit
• Gold, Colloidal
• Oil Red O
• Ponceau S
• Ponceau S Solution
• Reversible Protein Detection Kit for Membranes and Polyacrylamide Gels
• Stains–All
• Sudan Black B
• Fixing Solution, 5x concentrate
• CoZap for Coomassie® Blue Removal

Identify glycoproteins with superior selectivity using Sigma's new GlycoProfile III. Glycoprotein study, or Glycomics, is a rapidly growing, dynamic new field of research. Accurate initial identification of glycoproteins plays an important role in this research. Sigma understands and meets your needs with the most selective glycoprotein detection product available. ProteoProfile PTM Marker (Product Code P1745) contains a mix of glycosylated, phosphorylated and unmodified proteins and is available individually as well as with PP0300.

Superior Selectivity. ProteoProfile PTM Marker (P1745), containing glycosylated and non–glycosylated proteins, was separated by electrophoresis on a 4–20% SDS–PAGE gel. The gel was stained with GlycoProfile III (PP0300) (left), fluorescently imaged, and then stained for total protein with EZBlue™ Gel staining reagent (G1041) (right). Each band represents approximately 300 ng of protein.

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Alcian Blue 8GX is suitable for detection of glycoproteins on nitrocellulose and in PAGE gels. Alcian Blue 8GX reacts with sulfate and carboxylated functional groups. The interaction of Alcian Blue (cationic) and polyanionic glycoproteins is influenced by pH; both sulfate and carboxylate groups will react at pH 2.5, but sulfate groups alone react at pH 1.

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Amido Black Staining Solution is designed for rapid staining of protein bands on nitrocellulose membranes. Amido Black Staining Solution facilitates visualization of low concentration proteins with a low background. Proteins can be easily destained with 25% (v/v) isopropanol and 10% (v/v) acetic acid for further analysis. Amido black is also compatible with PAGE gels. Each bottle of 2X concentrate dilutes to 500 mL with water.

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Eosin Yellowish reversibly stains peptides and proteins dark red following SDS–PAGE. Proteins can then be recovered from the gel and further characterized.

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Fast Green FCF is used for protein detection and quantitation in native PAGE, SDS–PAGE, and IEF gels. Following staining, protein concentration can be measured at 625 nm. Fast Green staining is linear over a wider range of detection than Brilliant Blue R. It is especially well suited to IEF because it does not bind to ampholytes.

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This reagent reacts readily with primary amino groups in amino acids and peptides to form highly fluorescent compounds.

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The glycoprotein detection kit is designed to selectively stain glycoproteins in polyacrylamide gels and membranes using a modification of the Periodic Acid–Schiff (PAS) methods. Staining of sugar moieties of glycoproteins yields magenta bands with a colorless background. The detection limits using this kit are 25–100 ng of carbohydrate per band. Each kit contains sufficient materials for 10 mini–gels (10 x 10 cm).

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As a widely used protein and polysaccharide marker for electron microscopy, colloidal gold is a useful general stain for proteins immobilized on nitrocellulose membranes following PAGE.

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Oil Red O stain is suitable as a lipid/lipoprotein stain on cellulose acetate. A 2% stock solution in methanol is prepared and diluted with sodium hydroxide immediately before staining. The membrane is stained for approximately one hour and then destained in a methanol glycerol solution.

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Ponceau S (sodium salt) may be used to prepare a stain for rapid reversible detection of protein bands on nitrocellulose or PVDF membranes (Western blotting), as well as on cellulose acetate membranes. Common stain formulations include 0.1% (w/v) Ponceau S in 5% acetic acid or 2% (w/v) Ponceau S in 30% TCA and 30% sulfosalicylic acid.

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Ponceau S Solution is a ready–to–use, reversible staining solution designed for rapid (5 minute) staining of protein bands on nitrocellulose or PVDF membranes (Western blots), as well as cellulose acetate membranes. Ponceau S stain is easily reversed with water washes, facilitating subsequent immunological detection.

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The Reversible Protein Detection Kit for Membranes and Polyacrylamide Gels is a unique protein detection product for visualization of proteins on nylon, nitrocellulose, and PVDF membranes, as well as PAGE gels with sensitivity comparable to that of Coomassie stains. Many other stains produce high backgrounds on nylon membranes due to strong charge interactions. Application of activated R–PROB produces lavender stained protein bands, which are easily destained (reversed) with an EDTA solution. Each kit is sufficient for 200 applications (small blots on 8 x 10 cm PAGE gels).

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Stains–All is uniquely suitable for differential staining of nucleic acids and proteins. RNA bands appear bluish–purple, DNA bands appear blue, and proteins appear red. Typically, PAGE gels are stained in the dark and destained through exposure to light. Staining solutions are commonly made by dissolving the dye in formamide and buffer.

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Sudan Black B is a lipochrome (fat soluble dye) used to stain phospholipids, neutral fats, and sterols.

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Fixing solution prevents diffusion or loss of proteins prior to staining with a visualization reagent. 1x fixing solution contains 12% (w/v) trichloracetic acid and 3.5% (w/v) sulfosalicylic acid. Suitable for SDS–PAGE, non–denaturing PAGE, and IEF.

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Place the unique CoZap pad in your destaining tray and reduce destaining time 20%. CoZap absorbs any free dye in solution eliminating the need to change the destaining solution.

Product #	Product Name	Size
C4351	CoZap for Coomassie® Blue Removal	100 EA

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