Competition dialysis assay

Recently, Chairs analyzed the oxazolium pentamethine cyanine dye 35 (DODC, Fig. 14.8) in DNA binding studies utilizing his competitive dialysis method [75]. The nucleic acid structures included single-stranded, duplex, triplex, and tetraplex forms. The comparative assay demonstrated that DODC binds to triplex DNA more selectively than any of the tetraplex forms included in the assay. In addition, the... 

To evaluate the amount of acetaminophen bound to proteins, utilizing the competitive A-B ELISA or PCFIA, inhibition by unknown samples was compared with an assay standard prepared by derivatizing protein with NAPQI. For some experiments, 3-(N-acetyl-L-cystein-S-yDacetaminophen was used as an assay standard, in which case, the values obtained were corrected for differences in the relative inhibitory potency of 3-(N-acetyl-L-cystein- yl)acetaminophen and 3-Cys-A protein adduct (120 fmol/well and 2300 finol/well, respectively) (14). After dialysis, unknown samples were diluted to a final concentration of approximately 4 / g protein/assay well, assayed in duplicate, and expressed as nmoles of 3-Cys-A per mg of protein. The ELISA and PCFIA were shown to have similar limits of detection (20 pmole/mg protein) and to recognize the same itope as demonstrated by similar relative inhibitory potencies for N-acetylcysteine-acetaminophen, acetaminophen-bound... 

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