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DualColor™ Protein Loading Buffer Pack

#R1011 4X DualColor™ Protein Loading Buffer 20X Reducing Agent (2 M DTT)	for 1000 samples of 40 µl 2x5 ml 2x1 ml
Related Documents (in pdf, ~200 KB): Certificate of Analysis: #R1011 MSDS (English) MSDS (English-USA) MSDS (German) Flyer

 

Features

• Protects proteins from degradation during sample preparation for SDS-PAGE.
• Convenient - two dyes (blue and pink) allow for:
  • tracking electrophoresis progress,
  • monitoring Western blot protein transfer.

Description
The DualColor™ Protein Loading Buffer Pack is a complete solution for the preparation of protein samples prior to SDS-PAGE. Many proteins are sensitive to pH changes that result from temperature fluctuations during electrophoresis in Tris buffers. This optimized loading buffer prevents protein degradation during sample heating prior to SDS-PAGE (1) as well as during the electrophoresis run. The loading buffer contains two tracking dyes; blue (bromophenol blue) and pink (pyronin Y) which allow for tracking the progress of electrophoresis. In addition, the pink dye permits monitoring of protein transfer to the membrane during Western blot procedures. The DualColor™ Protein Loading Buffer Pack also contains SDS and DTT for complete disruption of all high-order protein structures. As DTT is prone to oxidation during multiple freeze-thaw cycles, it is supplied in separate vials.

Applications

• Preparation of proteins for SDS-polyacrylamide gel electrophoresis (SDS-PAGE).
• Monitoring protein transfer during Western blotting.

Composition

• 4X DualColor™ Protein Loading Buffer: 0.25 M Tris-HCl (pH 8.5 at 25°C), 8% SDS, 1.6 mM EDTA, 0.024% Pyronin Y, 0.04% Bromophenol blue and 40% glycerol.
• 20X Reducing Agent: 2 M DTT. 

Quality Control
Tested in protein samples preparation prior to SDS-PAGE and subsequent Western blotting.

Note
In 15% PAA gels pyronin Y dye migrates slower than bromophenol blue.

Storage
Store 4X DualColor™ Protein Loading Buffer at room temperature or at -20°C for long term storage.
Store 20X Reducing Agent at -20°C.

Figure 1. Increased protein stability with the DualColor™ Protein Loading Buffer.
12% Tris-glycine SDS-PAGE, proteins stained with PageBlue™ Protein Staining Solution.
M - PageRuler™ Unstained Protein Ladder
1, 3 - protein sample prepared with DualColor™ Protein Loading buffer (pH 8.5) and DTT
2, 4 - protein sample prepared with conventional Laemmli loading buffer (pH 6.8) and DTT

Figure 2. Visibility of the DualColor™ Protein Loading Buffer dyes.
12% Tris-glycine SDS-PAGE.
1-2 - protein samples prepared with DualColor™ Protein Loading Buffer Pack
3-4 - protein samples prepared with Protein Loading Buffer Pack

Related Products

ProteoJET™ Mammalian Cell Lysis Reagent ProteoJET™ Cytoplasmic and Nuclear Protein Extraction Kit Bradford reagent, ready-to-use Bovine Gamma Globulin Standard Set, ready-to-use Bovine Serum Albumin Standard Set, ready-to-use Protein Standard Solution PageRuler™ Plus Prestained Protein Ladder PageRuler™ Prestained Protein Ladder Prestained Protein Molecular Weight Marker PageRuler™ Unstained Protein Ladder Unstained Protein Molecular Weight Marker PageSilver™ Silver Staining Kit PageBlue™ Protein Staining Solution Protein Loading Buffer Pack 10X Tris-glycine-SDS 10X Tris-glycine 10X Tris-tricine-SDS DTT

Reference

1. Electrophoresis in practice, 4th edition, Westermeier, R., Wiley-VCH, 2005.

Updated vasario 27, 2008 11:41