[A new immunochemical method for detecting substance P receptors based on the biotin-streptavidin system].
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.
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry*; D001710:Biotin / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay; D007120:Immunochemistry; D008969:Molecular Sequence Data; D018040:Receptors, Neurokinin-1; D017981:Receptors, Neurotransmitter / Q000378:metabolism*; D019809:Streptavidin; D013373:Substance P / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity
D001426:Bacterial Proteins; D018040:Receptors, Neurokinin-1; D017981:Receptors, Neurotransmitter; D013373:Substance P; D001710:Biotin; D019809:Streptavidin
Novyĭ immunokhimicheskiĭ metod obnaruzheniia retseptorov veshchestva P na osnove biotin-streptavidinovoĭ sistemy.