Methods and compositions for detecting signals in binding assays
a technology of binding assay and signal detection, applied in the direction of microorganism testing/measurement, material analysis, biochemistry apparatus and processes, etc., can solve the problems of background noise, time and labor requirements, lack of specificity,
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[0074] A schematic of the current Affymetrix GeneChip® Expression Array signal amplification assay is shown below. Signal amplification over the signal obtained with the first streptavidin-phycoerythrin (SAPE) stain is achieved via a biotinylated anti-streptavidin antibody (b-Ab) that can complex with the SAPE from the first stain. The antibody contains multiple sites where multiple SAPE molecules can bind in the 3rd staining step.
[0075] Affymetrix GeneChip® Expression Array Signal Amplification:
[0076] probe:target-biotin=SAPE=biotinylated anti-streptavidin antibody (b-Ab)=SAPE [0077] step 1—complex biotinylated target:probe duplex with streptavidin-phycoerythrin (SAPE) [0078] step 2—complex biotinylated anti-streptavidin antibody to complex in step 1 [0079] step 3—complex multiple SAPE to complex in step 2
[0080] Microsphere particles were designed to be conjugated with the biotinylated anti-streptavidin antibody used in the current Affymetrix GeneChip® Expression Array signal am...
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