A new, very sensitive, bioluminescence-enhanced detection system for protein blotting. Ultrasensitive detection systems for protein blotting and DNA hybridization, I.
A relatively simple, very sensitive bioluminescence-enhanced detection system for protein blots is described. The method utilizes antibodies conjugated with alkaline phosphatase. The alkaline phosphatase then takes part in a reaction by releasing D-luciferin (Photinus pyralis) from D-luciferin-O-phosphate. Liberated D-luciferin reacts with luciferase, ATP and oxygen with light emission. The light is detected by a sensitive photographic film, thereby permitting the visualization of the alkaline phosphatase-conjugated antibodies. Under non-optimized conditions the limit of detection is at present 5 to 50 pg of protein, corresponding e.g. to 30 to 300 x 10(-18) mol of rabbit immunoglobulin G. The detection system is therefore 100 times more sensitive than other systems used at present.
Abteilung für Klinische Chemie und Klinische Biochemie in der Chirurgischen Klinik Innenstadt der Universität München.
R
R
Hauber
R
R
Geiger
D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D004247:DNA / Q000032:analysis*; D049454:Firefly Luciferin; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007118:Immunoassay; D008163:Luminescent Measurements; D009693:Nucleic Acid Hybridization; D011506:Proteins / Q000032:analysis*; D013552:Swine
D011506:Proteins; D049454:Firefly Luciferin; D004247:DNA; D000469:Alkaline Phosphatase