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  • TEMED-enhanced photoluminescent imaging of human serum proteins by quantum dots after PAGE.

TEMED-enhanced photoluminescent imaging of human serum proteins by quantum dots after PAGE.

Methods in molecular biology (Clifton, N.J.) (2012-05-16)
Na Na, Jin Ouyang
ABSTRACT

Polyacrylamide gel electrophoresis (PAGE) has been one of the most powerful and widely used separation techniques for complex biological samples, whose traditional detection methods include organic dye or silver staining. As a simple, convenient, and ultrasensitive detection of proteins for PAGE, a novel enhanced photoluminescent (PL) imaging method was developed. Thioglycolic acid (TGA)-capped CdTe quantum dots (QDs) and the enhanced reagent of tetramethylethylenediamine (TEMED) are introduced, achieving the direct detection of various proteins in native 1-DE, 2-DE, and SDS gels. Here, we describe the general protocol of TEMED-enhanced PL imaging by QDs, including materials, practical procedures, as well as some notes.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Brilliant Blue R, 250, suitable for microscopy
Sigma-Aldrich
Brilliant Blue R Concentrate, suitable for SDS-PAGE, methanol solution
Sigma-Aldrich
Brilliant Blue R, Dye content ~50 %, Technical grade
Sigma-Aldrich
Brilliant Blue R Staining Solution, suitable for (for immunoelectrophoresis protein staining)
Sigma-Aldrich
Brilliant Blue R, pure
Sigma-Aldrich
N,N,N′,N′-Tetramethylethylenediamine, ≥99.5%, purified by redistillation
Sigma-Aldrich
N,N,N′,N′-Tetramethylethylenediamine, ReagentPlus®, 99%
Sigma-Aldrich
N,N,N′,N′-Tetramethylethylenediamine, BioReagent, Molecular Biology, ≥99% (GC)
Sigma-Aldrich
N,N,N′,N′-Tetramethylethylenediamine, BioReagent, suitable for electrophoresis, ≥99.0%