Horseradish peroxidase -labeled

Y. Xiao, H.X. Ju, and H.Y. Chen, Hydrogen peroxide sensor based on horseradish peroxidase-labeled Au colloids immobilized on gold electrode surface by cysteamine monolayer. Anal. Chim. Acta. 391,... 

Detection reagent (substrate) for enzyme-labeled antibody 2,2-azino-di(3-ethyl-benzthiazohne sulfonate-6) (ABTS) 0.3 g/L in 0.15 M sodium citrate with 0.1% H2O2 for the detection of horseradish peroxidase-labeled secondary antibody or p-nitrophenyl phosphate (pNPP) 1 g/L in 1M diethanolamine in water for the detection of an alkaline phosphatase-labeled antibody (Kirkegaard and Perry Laboratories). 

Secondary antibodies are enzyme-linked antibodies directed against the primary antibody host animal s immunoglobulin (usually an IgG). If the primary antibody is mouse anticytokeratin the secondary antibody would be horseradish peroxidase-labeled or alkaline phosphatase-labeled antimouse IgG. 

Kricka, L. J., Stott, R A W., and Thorpe, G. H. G (1991) Enhanced chemiluminescent detection of horseradish peroxidase labels in ligand binder assays, in Luminescence Techniques in Chemical and Biochemical Analysis (Bayens, W R. G., De Kekeleire, D, and Korkidis, K, eds.), Dekker, New York, pp 599-635... 

Anti-atrazine antibodies were coupled to a Si microchip comprising 42 coated porous flow channels. The porous Si layer was achieved by anodizing the channel in 40% HF/96% ethanol (1 1). Detection of atrazine was achieved in a competitive format. The chemiluminescence signal decreased when a known amount of horseradish peroxidase-labeled atrazine was added. This decrease was used to determine the amount of unlabeled atrazine in the sample [1030],... 

Figure 25.1 Biotinylation of nanodiamond and recognition protocol with horseradish peroxidase-labeled streptavidin. Conditions and reagents (a) BH3 THF, THF, reflux, 72 h (b) APTMS, acetone, room temperature, 16 h (c) biotin, EDC, DMAP, CH2C12, 0°C — room temperature, 65 h.11...

Snitkoff et al. reported a specific immunoassay for monitoring the levels of ciprofloxacin in human samples [63], Serum was incubated at 37°C with shaking for 2 h, with rabbit anti-ciprofloxacin keyhole limpet haemocyanin primary antibody in microtiter wells coated with cipro-floxacin-BSA conjugate. Further incubation with horseradish peroxidase-labeled goat anti-rabbit IgG was carried out. The samples were shaken in 0.1 M citric buffer (pH 4.5), plates were incubated for 10 min, and then read at 450 nm. Calibration graphs were linear for 10 pg/mL to 10 ng/mL of ciprofloxacin, and the within-day RSD was less than 10%. 

Preston RJ, McCrea RA, Chang HT, Kitai ST (1981) Anatomy and physiology of substantia nigra and retrorubral neurons studied by extra- and intracellular recording and by horseradish peroxidase labelling. Neuroscience 6 331-344. 

A DNA optical sensor system was proposed by Cass and co-workers [35] based on the combination of sandwich solution hybridization, magnetic bead capture, flow injection and chemiluminescence for the rapid detection of DNA hybridization. Sandwich solution hybridization uses two sets of DNA probes, one labelled with biotin, the other with an enzyme marker and hybridization is performed in solution where the mobility is greater and the hybridization process is faster, rather than on a surface. The hybrids were bound to the streptavidin-coated magnetic beads through biotin-streptavidin binding reaction. A chemiluminescence fibre-optic biosensor for the detection of hybridization of horseradish peroxidase-labelled complementary DNA to covalent immobilized DNA probes was developed by Zhou and co-workers [36]. 

Horseradish, peroxidase-labeled goat, anti-mouse antibodies (Sigma-Aldrich Co., USA)... 

Chemiluminescence assays are ultrasensitive (attomole to zeptomole detection limits) and have wide dynamic ranges. They are now widely used in automated immunoassay and DNA probe assay systems, (e.g., acridinium ester and acri-dinium sulfonamide labels and 1,2-dioxetane substrates for alkaline phosphatase labels and the enhanced-luminol reaction for horseradish peroxidase labels [see Chapter 9]). 

Figure 9-14 Ultrasensitive assays for horseradish peroxidase and alkaline phosphatase labels. A, Chemiluminescent assay for horseradish peroxidase label using luminol. B, Chemiluminescent assay for an alkaline phosphatase label using AMPPD.

Ludwin SK, Kosek JG, Eng LE. The topographical distribution of SlOO protein and GEA protein in the adrrlt rat brain an im-mrmohistochemical study using horseradish peroxidase-labeled antibodies./ Comp Neurol. 1976 165 197-208. 

By utilizing horseradish-peroxidase-labeled OTA in western blots Schwerdt et al. [238] could show that in different renal cell lines (MEXTK-Cll, OK, LLC-PKj and immortalized human kidney epithelial cells (IHKE)) OTA binds directly to certain proteins. Thus OTA-protein binding damages normal protein function. Moreover, such binding results in a further accumulation of OTA [237]. 

Since the reports on enhanced chemiluminescence in the mid-1980s (e.g., TIO, T12-T14, T16, T17), numerous assays have been described in the literature. Table 5 represents a summary of the types of analyte that have more recently proved amenable to detection by enhanced luminol oxidation via horseradish peroxidase labels. This table is meant to be illustrative rather than comprehensive. 

Targeted drug delivery systems for the anticancer drug Taxotere were built from jS-CyD by monoconjugation with mannosyldendritic branches. Binding inhibition of horseradish peroxidase-labeled Concanavalin A to yeast mannan by mannosylated yS-CyDs was examined. The IC50 values of 95, 96, 97, 98, and 99 were 800, 780, 91, 110, and 8 pM, respectively. 97 solubilized Taxotere similarly to monobranched CyDs [34]. The solubility of Taxotere in water can by increased 1000-fold by 97. 

Fig. 1 Outline of the scheme for the production of horseradish peroxidase-labeled probes.

Hybridization of Horseradish Peroxidase-Labeled Probes and Detection by Enhanced Chemiluminescence... 

Horseradish peroxidase-labeled probe see Chapter 14) The labeling procedure produces a probe in a single-stranded form it should not be denatured further. 

SchOnhuber W, Zarda B, Eix S, Rippka R, Herdmann M, Ludwig W, Amann R (1999) In situ identification of cyanobacteria with horseradish peroxidase-labeled, rRNA-targeted oligonucleotide probes. Appl Environ Microbiol 65 1259-1267 Pernthaler A, Pernthaler J, Amann R (2002) Fluorescence in situ hybridization and catalyzed reporter deposition for the identification of marine bacteria. Appl Environ Microbiol 68 3094-3101... 

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