Molecular Weight Markers for Protein Electrophoresis

Protein molecular weight markers are used to calculate sample molecular weights, to monitor the progress of an electrophoretic run, or as a positive control for analysis conditions. Sigma offers a wide selection of markers for numerous protein electrophoresis applications, including silver staining, isoelectric focusing, fluorescent studies, and many more. All have been use-tested to assure outstanding performance and are available in convenient package sizes.

Prestained Markers
• ColorBurst™ Markers
• Color Markers
• Prestained Blue Marker

Unstained Markers
• SigmaMarkers™
• Ultra-Low Range Marker
• Peptide Molecular Weight Marker
• Other Unstained Molecular Weight Markers

Specialty Markers
• Chemichrome™ Western Control
• Chemichrome™ Ultimate
• ProteoProfile™ PTM Marker
• Biotinylated Markers
• 2D Electrophoresis Marker
• Fluorescent Markers
• Recombinant Markers
• Silver Stain Markers
• SYPRO® Marker
• Non-Denaturing Molecular Weight Marker
• IEF Marker (3.6-9.3)

Brilliantly colored, exceptionally well-resolved, convenient, and stable, ColorBurst protein molecular weight marker performs impressively in a variety of gel compositions and concentrations. ColorBurst is composed of eight proteins ranging from 8 to 220 kDa, which have been chemically reduced, alkylated, and conjugated to brilliantly colored dyes. ColorBurst can be used to estimate sample molecular weights, to monitor the progress of an electrophoretic run, or to confirm that an electroblot is complete. Each vial contains 500 ml of marker solution and may be used for up to 100 applications.

Features & Benefits

• Exceptional resolution allows estimation of sample molecular weights
• No freeze / thaw cycle means decreased degradation and long shelf life
• Ready-to-use No boiling required before use
• No chemical reduction necessary prior to gel loading

C1992 A 10-20% Tris-Glycine gel was loaded (left to right) with 3, 5 and 7 µl of the ColorBurst Marker. The markers were run using standard conditions on 10 x 10 cm, 1 mm thick, 10-well precast gels. Bands transferred to nitrocellulose membranes from the gels in Figure 1 (Tris-Glycine on left and Tris-Tricine on right). Transfers were completed in 90 minutes at 70 volts with Towbin's buffer (Tris-Glycine in 20% methanol.)

C4861 The gel was loaded (left to right) with 3, 5, and 7 µl of ColorBurst Marker. The marker was run using standard conditions on a 10 x 10 cm, 1 mm thick, 10 well precast 4-20% Tris-Glycine gel. Bands were transferred to nitrocellulose membrane. The transfer was completed in 90 minutes at 70 volts using Towbin’s buffer (Tris-Glycine in 20% methanol).

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Sigma Color Markers contain 5-6 proteins conjugated to colored dies providing a monitor for protein migration and transfer efficiency. These markers are available in low range, suitable for use in the Laemmli SDS-PAGE system and ultra-low range for use in a Tricine SDS-PAGE system.

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Sigma’s prestained blue consists of 7 proteins conjugated to a blue dye. This marker provides a monitor for protein migration and transfer efficiency.

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SigmaMarkers encompass the range of molecular weights common to most proteins and their subunits. Lyophilized with sample buffer, the wide, high, and low range markers are ready for use following reconstitution with water. They are formulated to yield a distribution of well- defined bands of approximately equal intensity following electrophoresis and Coomassie® Blue staining.

5 µl of SigmaMarkers S8445, M3913 and S8320 were run on a 4-20% PAGE gel and stained with EZBlue (G1041)

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Sigma Molecular Weight Markers are available in ultra-low range weight ranges. The ultra-low range marker is suitable for use in a Tricine SDS-PAGE system.

5 µl of a 1:20 dilution of the Ultra-low Range Molecular Weight Marker (M3546) was run on a 10-20% Tris-Tricine SDS-PAGE gel, fixed with 5% gluteraldehyde and stained with EZBlue (G1041).

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The Peptide Molecular Weight Marker is a mixture of seven polypeptides ranging for 2,500 to 17,000 Da.

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Sigma’s long standing markers, SDS7 and SDS6H2 previously titled Dalton markers, contain a mixture of proteins suitable for use in both the Weber Osborn (1) Electrophoresis System and the Laemmmli SDS-PAGE system.

1. Weber, K.; Osborn, M. J. Biol. Chem. 244:4406, (1969)

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Chemichrome Western Control is an ideal molecular weight marker and positive control for electrophoresis and subsequent Western blotting. Similar to ColorBurst™, Chemichrome contains an additional band of mouse IgG. During electrophoresis, brightly colored protein bands serve as positive controls for protein migration. During blotting, the same brightly colored bands indicate that the transfer is complete. After incubation with mouse primary and secondary antibodies, the band of mouse IgG (heavy chain) serves as a positive control using either colorimetric or chemiluminescent substrates. Simple to use and extremely practical, Chemichrome Western Control enables researchers to confirm the success of their analyses every step of the way. Chemichrome is supplied in vials containing 200 µl of ready-to-use solution.

Features & Benefits

• Confirms that a membrane transfer is complete
• Confirms successful Western Blotting conditions
• Compatible with many peroxidase and phosphatase substrates, including TMB and ECL®

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Like Colorburst™ and Chemichrome™ Western Control, Chemichrome Ultimate contains eight brilliantly colored polypeptides which serve as positive controls during protein migration and transfer. In addition to mouse IgG, Chemichrome Ultimate also contains goat and rabbit IgG, which serve as positive controls for Western blotting using either colorimetric or chemiluminescent substrates.

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Glycosylation and phosphorylation are frequently encountered post-translation modifications in proteomic analysis. ProteoProfile PTM Marker, containing glycosylated and phosphorylated proteins, is designed for use as a positive and negative control in studying these post- translational modifications of proteins in SDS-PAGE. Electrophoresis will yield four protein bands:

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. BSA (not glycosylated, not phosphorylated) Ovalbumin (glycosylated and phosphorylated) b-Casein (not glycosylated, phosphorylated) RNase B (glycosylated, not phosphorylated)

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Biotinylated molecular weight markers contain biotin-conjugated proteins which, following electrophoresis and membrane transfer, can be easily detected with a Streptavidin-Peroxidase Conjugate (S5512) and a color development reagent such as TMB (T0565).

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Specifically designed for denaturing IEF and 2D electrophoresis, this molecular weight marker set contains four proteins selected to provide a diagonal pattern on a wide range 2D gel.

5 µl of 2D markers (M3411) were separated with a mixture of ampholine pH 3.5-9.5 and ampholine pH 2.5-4.5 in the ratio 1:2.5 and then separated on a 12.5% SDS-PAGE gel. The gel was stained with Coomassie Brilliant Blue G (B8522). Amyloglucosidase from Aspergillus niger (A8437) Ovalbumin (O4757) Carbonic Anhydrase from Human Erythrocytes (C4806) Myoglobin from Horse Heart (M7911)

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Sigma fluorescent molecular weight markers are composed of FITC-conjugated proteins, which are well resolved and easily visualized with ultraviolet light in polyacrylamide gels or membranes.

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The Recombinant Molecular Weight Standard Mixture contains seven precisely sized proteins, molecular weights 15, 25, 35, 50, 75, 100, and 150 kDa. Recombinant proteins form sharp bands for accurate sample MW calculations.

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Designed for molecular weight determinations on silver stained gels, Silver Stain SDS- PAGE molecular weight standard mixtures contain evenly distributed, well resolved proteins of equal intensity. Check out our ProteoSilver™ Stains.

7 µl of a 1:15 dilution of the Silver Stain Molecular Weight Marker (M6539) was run on a 10-18% SDS-PAGE gel and silver-stained.

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The SYPRO marker contains 11 proteins suitable for use in SDS-PAGE. Detection is completed with a SYPRO fluorescent protein gel stain.

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Sigma's Non-Denaturing Marker contains 5 separate proteins suitable for use in a native polyacrylamide gel electrophoresis system.

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The IEF Marker 3.6-9.3 is a mixture of proteins with well-defined pI values. The protein mixture forms evenly spaced, distinct bands over the designated pH range.

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