Sensitive quantification of proteins by electrophoresis in gels by use of chemiluminescence.
For protein quantitation in gels or blotting membranes, chemiluminescence (CL) offers the advantages of a substantial improvement of detection limits. Easy-to-use CL-chemicals and specific probes such as antibodies conjugated to enzymes, e.g. alkaline phosphatase (AP) may be used in combination with a newly developed luminometer. CL was found to have low detection limits and a linear relation between relative light units (RLU) and the concentration of the antibody enzyme complex present over a wide concentration range. Measurements of the immunoglobulin IgE in dot blots and in blots after sodium dodecyl sulfate (SDS) electrophoresis under nonreducing conditions in agarose gels are described.
Division of Respiratory Allergology and Immunology, IMA, National Institute of Occupational Health, Solna, Sweden.
O
O
Vesterberg
F
F
Acevedo
C
C
Bayard
D000469:Alkaline Phosphatase; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D007124:Immunoenzyme Techniques; D007073:Immunoglobulin E / Q000032:analysis*; D008163:Luminescent Measurements; D008567:Membranes, Artificial; D011145:Polyvinyls; D012967:Sodium Dodecyl Sulfate; D005465:Fluorocarbon Polymers
D000469:Alkaline Phosphatase; D007073:Immunoglobulin E; D008567:Membranes, Artificial; D011145:Polyvinyls; D012967:Sodium Dodecyl Sulfate; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers