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| | HTRF Biochemical Assay |
| Description: | Among the advantages of this Abeta 42 HTRF gamma-secretase assay is the construction of the biotinylated, recombinant CT6-I45F substrate, which has provided a highly active substrate allowing for increased sensitivity and detection of gamma-secretase activity. This recombinant protein is based on the sequence of APP, but has a truncation that removes an auto-inhibitory domain. This provides markedly increased activity. Furthermore, an AviTag site has been cloned into the vector encoding this substrate. Consequently, overproduction of the CT6-I45F substrate in the presence of biotin ligase and biotin results in direct biotinylation of our substrate. This novel approach further increases the sensitivity of the assay. Other attempts at development of a high throughput gamma-secretase assay that screens for Abeta 42 have been unsuccessful. This is the first successful development of a biochemical assay that screens for gamma-secretase cleavage at the Abeta 42-site. |
| Affinity data for this assay | |
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