Polymerization of acrylamide at acid pH using uranyl nitrate

Area of Science:

• Biochemistry
• Polymer Chemistry
• Analytical Chemistry

Background:

• Traditional acrylamide gel polymerization often uses persulfate, requiring high concentrations and pre-electrophoresis for removal.
• Low pH polymerization presents challenges for protein separation and gel reproducibility.

Purpose of the Study:

• To introduce uranyl nitrate as a novel photopolymerizing reagent for acrylamide gels at low pH.
• To evaluate the efficiency and effectiveness of uranyl nitrate compared to persulfate in gel polymerization and protein electrophoresis.

Main Methods:

• Polymerization of acrylamide gels using uranyl nitrate at pH 3.0.
• Comparison of uranyl nitrate and persulfate concentrations for gel polymerization.
• Electrophoretic separation of basic proteins using gels polymerized with uranyl nitrate.
• Assessment of enzymatic activity of trypsin after electrophoresis in uranium-polymerized gels.

Main Results:

• Uranyl nitrate effectively polymerizes acrylamide gels at low pH (3.0) using significantly lower concentrations (0.2 mg/ml) than persulfate (7 mg/ml).
• Gels polymerized with uranyl nitrate demonstrate enhanced protein separation, faster movement, and better resolution for basic proteins.
• The use of uranyl nitrate eliminates the need for pre-electrophoresis to remove excess initiator.
• Electrophoresis in uranium-polymerized gels does not compromise the enzymatic activity of proteins like trypsin.
• The reagent is also effective for polymerizing gels containing 3 M urea.

Conclusions:

• Uranyl nitrate is a highly efficient and versatile photopolymerizing reagent for acrylamide gel electrophoresis at low pH.
• This method offers advantages in terms of reduced reagent cost, simplified procedures, and improved electrophoretic performance.
• Uranium-polymerized gels are reproducible and suitable for sensitive applications, including the analysis of enzyme activity.