A new luminometer for sensitive quantification by chemiluminescence of specific proteins in microtitre plates and on blot membranes.
Generally applicable technologies are described, which depend on the use of chemiluminescence and a new type of a versatile luminometer, that can measure also weak light in microtitre plates (microplates) and on blotting membranes that is especially useful for dot blot immunoassay. Applications are described using alkaline phosphatase conjugated antibodies against IgG and IgE and the reagent Lumi-Phos 530. This chemiluminescence offers advantages over the use of radioactive isotopes, densitometry and light reflection measurement on membranes and also ELISA, for sensitive quantification of e.g. specific proteins. Special procedures are described for the first time that with the mentioned reagent in agarose gel allows specific and very sensitive quantification of proteins on dot blot membranes. The luminometer, which has temperature control, is very sensitive, precise and allows efficient protocols for various assays. It thus fulfils many of the requirements for good quantification, is time-saving and in addition brings significant improvements due to very low detection limits and large linear concentration ranges that can be measured with excellent regression coefficients (r2 often about 0.999).
Vesterberg
O
O
Division of Respiratory Allergy and Immunology, IMA, National Institute of Occupational Health, Solna, Sweden.
D000469:Alkaline Phosphatase / Q000032:analysis*; D000818:Animals; D003720:Densitometry / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006041:Goats; D006801:Humans; D007073:Immunoglobulin E; D007074:Immunoglobulin G; D008027:Light; D008163:Luminescent Measurements; D010783:Photometry / Q000295:instrumentation* / Q000379:methods; D011506:Proteins / Q000032:analysis*; D011817:Rabbits; D011865:Radioisotope Dilution Technique; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D007074:Immunoglobulin G; D011506:Proteins; D007073:Immunoglobulin E; D000469:Alkaline Phosphatase