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mPAGE® Bis-Tris Precast Gels for Gel Electrophoresis

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Avoid breaking the bank for publication-ready, gel electrophoresis images with mPAGE® bis-tris precast gels. Compatible with most electrophoresis tanks, mPAGE® precast gels can be used to achieve high-resolution protein separation with a shorter run time (up to 15 minutes).
  • High-resolution separation
  • Easy-to-use with common electrophoresis tanks
  • Increased sample well volume (up to 80 μL)
  • Compatible with wet and semi-dry Western blot transfer
  • Shorter run time (up to 15 minutes)
  • Prevents protein modification with neutral pH

Protein separation and transfer with mPAGE® gels

Polyacrylamide gel electrophoresis (PAGE) is a common technique used to separate proteins. While folded (or native) proteins can be separated using native-PAGE, proteins are typically unfolded with sodium dodecyl sulfate (SDS) and a reducing agent like dithiothreitol (DTT) prior to electrophoresis. As SDS-PAGE gels separate proteins by their molecular weight, samples are compared to standards (ladders) with proteins of known molecular weight for analysis.

Separation

Depending on the gel and conditions, gel electrophoresis run time can be highly variable. In comparison to other commercially available precast PAGE gels, mPAGE® bis-tris precast gels were shown to have up to a 15-minute shorter run time (Figure 1), without resolution loss.

Competitor T
50-minute run time		 mPAGE® Precast Gel
35-minute run time
Comparison of the separation of a serial dilution of E. Coli extract on a mPAGE<sup>®</sup> precast gel and a commercially available alternative. The mPAGE gel provided identical resolution in only 35 minutes, compared to the alternative product that requires 50 minutes for the same run. Figure shows the images of two stained, Western blotting membranes side-by-side after immunodetection. Both membranes show the protein separation achieved when performing gel electrophoresis on an E. Coli extract with a precast polyacrylamide gel in MES running buffer. The left membrane image was generated using a competitor’s precast PAGE gel, while the right membrane image was generated using a Millipore mPAGE<sup>®</sup> precast PAGE gel. Both membrane images show identical resolution for protein separation.
Figure 1. Protein separation using MOPS running buffer comparing mPAGE® Bis-Tris Precast Gel (4-12%) to a competitor precast bis-tris gel. Samples: unstained protein marker and serial dilution of E.coli extract.

Transfer to membranes

After separation, proteins are often transferred to membranes for detection, in a technique known as Western blotting (immunoblotting). In Western blotting, proteins are transferred using either wet or semi-dry electrophoresis transfer methods. mPAGE® bis-tris precast gels provide efficient transfer for both electrophoresis methods (Figure 2).

Wet Transfer
Wet (tank) transfer and semi-dry transfer are the two most common transfer techniques used in Western blotting protocols. mPAGE precast gels provide good transfer for both methods. The image shows quality protein transfer from a serially diluted sample of A431 cell lystate for both wet and semi-dry transfer.

Semi-dry Transfer
Figure shows the stacked images of two Western blotting membranes after the immunodetection step. The proteins in the top image were transferred to the membrane using a wet transfer electrophoresis method. The proteins in the bottom image were transferred to the membrane using a semi-dry transfer electrophoresis method. Both images look identical, verifying that separated proteins can be transferred from mPAGE<sup>®</sup> gels using either wet or semi-dry transfer methods.
Figure 2. Serial dilutions of A431 cell lysate loaded onto an mPAGE® 4-12% gel and transferred to Immobilon®-P membrane via standard methods. Blots probed with antibodies against EGFR (180 kDa), AKT1 (60 kDa) and histone (17 kDa). Antibodies detected with peroxidase-conjugated secondary antibodies and Immobilon® Forte chemiluminescence substrate.

How to select mPAGE® precast gels

Precast gels are available in different gel concentrations (percentages) and are selected based on the protein size. In addition to different percentages, precast gels are either available at fixed acrylamide percentages or as gradient gels where the concentration of acrylamide varies throughout the gel. Low percentage gels are typically used to separate high molecular weight proteins, while high percentage gels are used to separate low molecular weight proteins. Gradient gels are typically used to separate broad protein ranges, or mixtures including both high and low molecular weight proteins. The choice of MOPS or MES running buffer can be used for further tuning of the separation. MOPS can provide improved separation at the higher range of the gel, and MES can improve separation at the lower range of the gel.The banding patterns in the migration charts below in MOPS (Figure 3) and MES buffer (Figure 4) can be used to identify the correct buffer and mPAGE® precast gel for your application.

Protein banding patterns for different mPAGE<sup>®</sup> gel strengths run in Tris-MOPS running buffer. This migration chart can be used to determine the best mPAGE<sup>®</sup> gel concentration and format for your gel electrophoresis application." title="Protein banding patterns for different mPAGE<sup>®</sup> gel strengths run in Tris-MOPS running buffer. This migration chart can be used to determine the best mPAGE<sup>®</sup> gel concentration and format for your gel electrophoresis application.

Figure 3. Migration chart for mPAGE® precast gels using Tris-MOPS SDS running buffer


Protein banding patterns for different gel mPAGE<sup>®</sup> strengths in MES SES running buffer. This migration chart can be used to determine the best mPAGE<sup>®</sup> gel concentration and format for your gel electrophoresis application." title="Protein banding patterns for different gel mPAGE<sup>®</sup> strengths in MES SES running buffer. This migration chart can be used to determine the best mPAGE<sup>®</sup> gel concentration and format for your gel electrophoresis application.

 Figure 4. Migration chart for mPAGE® precast gels using MES SDS running buffer
 

mPAGE® Gels, Buffers, and Accessories

 
mPAGE® Precast Gels
DescriptionQuantityCatalog No.
mPAGE® 4-12% Bis-Tris, 10x8, 10 well 10 gels MP41G10
mPAGE® 4-12% Bis-Tris, 10×8, 12 well 10 gels MP41G12
mPAGE® 4-12% Bis-Tris, 10×8, 15 well 10 gels MP41G15
mPAGE 4-20% Bis-Tris, 10x8, 10-well 10 gels MP42G10
mPAGE 4-20% Bis-Tris, 10x8, 12-well 10 gels MP42G12
mPAGE 4-20% Bis-Tris, 10x8, 15-well 10 gels MP42G15
mPAGE 8-16% Bis-Tris, 10x8, 10-well 10 gels MP81G10
mPAGE 8-16% Bis-Tris, 10x8, 12-well 10 gels MP81G12
mPAGE 8-16% Bis-Tris, 10x8, 15-well 10 gels MP81G15
mPAGE 8% Bis-Tris, 10x8, 10-well 10 gels MP8W10
mPAGE 8% Bis-Tris, 10x8, 12-well 10 gels MP8W12
mPAGE 8% Bis-Tris, 10x8, 15-well 10 gels MP8W15
mPAGE 10% Bis-Tris, 10x8, 10-well 10 gels MP10W10
mPAGE 10% Bis-Tris, 10x8, 12-well 10 gels MP10W12
mPAGE 10% Bis-Tris, 10x8, 15-well 10 gels MP10W15
mPAGE 12% Bis-Tris, 10x8, 10-well 10 gels MP12W10
mPAGE 12% Bis-Tris, 10x8, 12-well 10 gels MP12W12
mPAGE 12% Bis-Tris, 10x8, 15-well 10 gels MP12W15
 
mPAGE® Buffers
DescriptionQuantityCatalog No.
mPAGE 4X LDS Sample Buffer 10 mL MPSB-10ML
mPAGE 4X LDS Sample Buffer 250 mL MPSB-250ML
mPAGE MES SDS Running Buffer Powder 5 packets (1L/packet) MPMES
mPAGE MOPS SDS Running Buffer Powder 5 packets (1L/packet) MPM0PS
mPAGE Transfer Buffer Powder 10 packets (1L/packet) MPTRB
 
mPAGE® Trial Packs
DescriptionContentsCatalog No.
mPAGE Trial Kit, 4-12%, 12-well, MOPS
2 gels, 1L Running Buffer Powder, Adapter Plates, Cassette Opener
 MP41G12TR1
mPAGE Trial Kit, 4-20%, 12-well, MOPS MP42G12TR1
mPAGE Trial Kit, 10%, 12-well, MOPS MP10W10TR1
mPAGE Trial Kit, 4-12%, 12-well, MES MP41G12TR2
mPAGE Trial Kit, 4-20%, 12-well, MES MP42G12TR2
mPAGE Trial Kit, 12%, 12-well, MES MP12W12TR2
 
mPAGE® Accessories
DescriptionQuantityCatalog No.
mPAGE Adapter Plates 2 MPTA
mPAGE Gel Cassette Opener 1 MPC0
mPAGE Buffer Dam 1 MPBD