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.
Vesterberg
O
O
Division of Respiratory Allergology and Immunology, IMA, National Institute of Occupational Health, Solna, Sweden.
Acevedo
F
F
Bayard
C
C
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