Catalyzed reporter deposition

Tyramide signal amplification This procedure, designated as a catalyzed reporter deposition (CARD) or tyramide signal amplification (TSA), takes advantage of horseradish peroxidase (HRP) from an HRP-labeled secondary antibody to catalyze in the presence of hydrogen peroxide the oxidation of the phenol moiety of labeled tyramine. On oxidation by HRP, activated tyramine molecules rapidly bind covalently to electron-rich amino acids of proteins immediately surrounding the site of the immunoreaction. This allows an increase in the detection of an antigenic site up to 100-fold compared with the conventional indirect method with no loss in resolution. 

Hunyady, B Krempels, K., Harta, G. Y and Mezey, E. (1996) Immunohis-tochemical signal amplification by catalyzed reporter deposition and its application in double immunostaining. J. Histochem. Cytochem. 44, 1353-1362. 

In immunocytochemistry, signals are usually amplified by means of catalyzed reporter deposition. In ProxiMol, catalyzed reporter deposition has been modified to permit the isolation of phage antibodies binding to near neighbor biotinylated guide molecule. Deposition of biotin molecule can be detected by electron microscopy. 

Different authors and commercial suppliers have assigned different names to signal amplification using tyramine. For example, tyramine signal amplification (TSA) system and the catalyzed signal amplification (CSA) system are comma-daily available from DuPont NEN Life Science Products, Boston, MA, and DAKO Corporation, Carpinteria, CA, respectively. In addition, the tarns CARD (catalyzed reporter deposition) (Bobrow et al., 1989), TA (tyramide amplification) (Shindler and Roth, 1996), and ImmunoMax (Merz et al., 1995) have been used for the tyramine amplification technique. The use of different names for almost identical procedures has resulted in confusion. To standardize the terminology, the neutral abbreviation, tyramide amplification technique (TAT) should be accepted (Von Wasielewski et al., 1997). 

Compared with radioactive ISH, nonradioactive ISH requires a 10- to 50-fold higher concentration of probes such as oligonucleotides. However, signal amplification is decreased by increasing probe concentration. Therefore, since nonradioactive probes have limited sensitivity, especially when applied to low-abundance mRNAs, a technique is required for signal amplification. One such technique consists of an optimized protocol for rapid signal amplification based on catalyzed reporter deposition (CARD) that increases the sensitivity of nonradioactive mRNA ISH on the formaldehyde-fixed and paraffin-embedded tissues (Speel et al., 1998). This technique facilitates the detection of low-copy mRNAs by ISH (Yang et al., 1999). 

Bobrow, M. N., Harris, T. D., Shaughnessy, K. J., and Litt, G. J. 1989. Catalyzed reporter deposition, a novel method of signal amplification. Application to immunoassays. J. Immunol. Methods 725 279-285. 

Totos, G., Thakhi, A., Hauser-Kronberger, C., and Tubbs, R. R. 1997. Catalyzed reporter deposition a new era in molecular and immunomorphology-nanogold-silver staining and colorernctric detection and protocols. Cell Vis. 4 433-437. 

Wigle, D. A., Radakovic, N. N., Venance, S. L., and Pang, S. C. 1993. Enhanced chemiluminescence with catalyzed reporter deposition for increasing the sensitivity of western blotting. Biotechniques 74 562-563. 

Gold labeling may be used in the recently introduced catalyzed reporter deposition-immunogold technique where biotinylated tyramide molecules are attached the antibody-antigen complex site the biotinylated sites are visualized by interaction with streptavidin-gold (46,47). 

Pernthaler, A., Pemthaler, J., and Amann, R. (2002a). Fluorescence in situ hybridization and catalyzed reporter deposition for the identification of marine bacteria. Appl. Environ. Microbiol. 68, 3094-3101. 

Behrens, S., Losekann, T., Pett-Padge, J., Weber, P. K., Ng, W. O., Stevenson, B. S., et al. (2008). Linking microbial phylogeny to metabolic activity at the single-cell level by using enhanced element labeHng-catalyzed reporter deposition fluorescence in situ hybridization (EL-FISH) and NanoSIMS. Appl. Environ. Microbiol. 74, 3143—3150. 

Fig. 7.8. Amplification generated by the production of a catalytically active NAD. The main problem with this technique is that even trace amounts of NAD in the enzyme or NADP preparations, will lead to high backgrounds. Alternative amplification methods include enzyme cascades (Blake et al., 1984) and catalyzed reporter deposition (Bobrow et al., 1991).

Based on the principle of enzyme amplification for enzyme immunoassays adopted in the 1980s, Bobrow and associates developed a catalyzed reporter deposition technique (CARD) to achieve amplified signal for solid-phase immunoassay system and membrane immunoassays. Subsequently, this CARD technique was introduced to IHC in 1992. ... 

PAP, peroxidase-antiperoxidase ABC, avidin-biotin conjugate APAAP, alkaline phosphatase antialkaline phosphatase B-SA, biotin-streptavidin PCNA, proiiferating cell nuclear antigen EPOS, enhanced polymer one-step staining (Dako) CARD, catalyzed reporter deposition HRP, horseradish peroxidase. 

Bobrow MN, Harris TD, Shaughnessy KJ, et al. Catalyzed reporter deposition A novel method of signal amplification Application to immunoassays. J Immunol Methods. 1989 125 279. 

Bobrow MN, Litt GJ, Shaughnessy KJ, Mayer PC, Conlon J (1992) The use of catalyzed reporter deposition as a means of signal amplification in a variety of formats. J Immunol Meth 150 145-149... 

Figure 3.11 Reverse-phase microarray. In the streptavidin linked to reporter molecules such RPMA, proteins extracted from cellular lysates as QDs or enzymes such as HRP that carry are arrayed onto a nitrocellulose substrate out catalyzed reporter deposition for signal and probed with a primary antibody. In turn, a amplification. (Reprinted with permission biotinylated secondary antibody recognizes from Reference [51]. Copyright 2007 American the presence of the primary antibody. The Chemical Society.) biotinyl groups are then detected by...

CARD-FISH catalyzed reporter deposition-fluorescence hybridization... 

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