Chromogenic substrates for horseradish peroxidase.
Two new detection systems for horseradish peroxidase (HRP) have been developed for the staining of membranes used in immunoassays. These systems use dimethyl or diethyl analogues of p-phenylenediamine with 4-chloro-1-naphthol to generate a blue product or 3-methyl-2-benzothiazolinone hydrazone with 4-chloro-1-naphthol to generate a red product. These reagents offer increased sensitivity and lower background staining than currently available chromogenic detection substrates. In addition, the incorporation of these substrates increases the sensitivity of HRP labels to be comparable to that of alkaline phosphatase with the 5-bromo-4-chloro-3-indolyl phosphate + nitro blue tetrazolium substrate.
Conyers
S M
SM
Chemistry Division, Naval Research Laboratory, Washington, D.C. 20375-5000.
Kidwell
D A
DA
D000469:Alkaline Phosphatase / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology; D002863:Chromogenic Compounds / Q000378:metabolism*; D004396:Coloring Agents / Q000378:metabolism*; D006735:Horseradish Peroxidase / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000378:metabolism; D012680:Sensitivity and Specificity; D013379:Substrate Specificity
D002863:Chromogenic Compounds; D004396:Coloring Agents; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase; D000469:Alkaline Phosphatase