Signal reagent

Application of Novel Acridan Esters as Chemiluminogenic Signal Reagents in Immunoassay... 

In 1994 at the 8th International Symposium on Bioluminescence and Chemiluminescence, Schaap and Akhavan-Tafti presented a new technology for the detection of HRP [13, 19, 20], They proposed the use of certain aromatic acridan esters (see Fig. 4 for the structures) as part of a signal reagent for very sensitive detection... 

Although the signal reagent can be used over a wide pH range, the optimal pH was found to be 5-7. Most of the results described in this chapter were obtained using a pH of 5.4 [phosphate-buffered saline (PBS), 10 mM],... 

As can be seen from Figure 7, the signal reaches a plateau within 10 min and lasts for a relatively long time (even after 5 h a useful signal can be obtained). Furthermore, the signal reagent allows measurement of HRP over a wide range... 

Figure 8 HRP calibration curve (log-log plot of HRP dose vs. chemiluminescence signal, corrected for background) using signal reagent incorporating GZ-11.

To test our new signal reagent based on GZ-11 the detection system was applied to two competitive-format immunoassays. These two assays (for atrazine and clenbuterol) normally use chromogenic detection systems. While these colorimetric assays may be adequate for laboratory use, chemiluminescent detection offers potential advantages in sensitivity and on site screening applications [33],... 

In practical terms, microarrays are operational by at least two means. One can (1) create very small spots of capture ligands that bind only appreciable amounts of analyte and measure binding with ultrasensitive signaling reagents and detectors or (2) create arrays of much larger spots of capture ligands that significantly deplete the sample of analyte and permit the use of less sensitive (and presumably less expensive) approaches to detection. 

Amerlite signal reagent (Johnson and Johnson Clinical Diagnostics) is supplied as separate bottles of buffer and substrate tablets. One A and one B substrate tablet (cat. no. LAN.4401) are dissolved in each bottle of substrate buffer (cat no. LAN. 4402) prior to use. This reagent is stable for a day at room temperature providing it is kept in the dark glass bottle... 

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