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  • Title: [A new immunochemical method for detecting substance P receptors based on the biotin-streptavidin system].
    Author: Golovnina TA, Kasheverov IE, Plaksin DIu, Iakunina NB, Kovalenko VA, Utkin IuN, Tsetlin VI.
    Journal: Bioorg Khim; 1992 Jul; 18(7):932-41. PubMed ID: 1280142.
    Abstract:
    A novel method for detecting the membrane receptors of Substance P (SP) has been developed. The method does not require radioactive derivatives of SP and is based on quantitation of specifically bound biotinylated SP (Bt-SP), the analysis being performed after destruction of the ligand-receptor complex in acidic conditions and Bt-SP extraction into solution. The acidic extract from the Bt-SP-membrane complex after neutralization is added to immulon microELISA plate coated with affinity purified anti-SP-antibodies. The sensitivity of subsequent detection of Bt-SP enhances by using a new type of the streptavidin conjugate with a polymeric form of horseradish peroxidase. Enzyme immunoassay conditions were optimized using [125I]-labelled SP derivative. The developed method allows the determination of femtomoles of SP in a sample and has a sensitivity comparable to that of radioligand analysis.
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