CDP-Star™ as a Chemiluminescent Substrate for Use with Alkaline Phosphatase Labelled Probes
In the last five years alkaline phosphatase-labelled probes have been widely adopted for use in DNA fingerprinting and DNA profiling eg NICE™ probes (Cellmark) (Ref. 1). Apart from the obvious operational advantages of using non-radioactive systems, hybridisation with alkaline phosphatase-labelled probes is far more rapid than with traditional systems. Typically membranes can be hybridised and autoradiographs produced within seven hours. Hybridisation is detected by the application of a chemiluminescent substrate to the membrane. The substrate is activated by the alkaline phosphatase label on the probe resulting in emission of visible light which is recorded on X-ray film. One of the slowest parts of the procedure is exposure of hybridised membranes to X-ray film, which takes 3–4 hours or more. In an effort to shorten exposures and reduce the overall length of the DNA profiling process, we examined the performance of two different chemiluminescent substrates, Lumi-Phos® 530 (Lumigen Ine, Ref. 2) and CDP-Star™ (Tropix Inc, Ref. 3) which is reported to give shorter exposure times.
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