Revvity, 64APBKAA, HTRF Human APP (BACE1 Substrate) Biotin Eu-Conjugate, 5,000 Assay Points

Beta-secretase (BACE1) is an enzyme which cleaves amyloid precursor protein (APP) to produce the amyloid ß42 peptide, responsible for insoluble amyloid plaques in Alzheimer's disease (AD). HTRF APP substrate labeled with both biotin and Eu3+ Cryptate, used with HTRF SA-XL665 or Sa-d2, provides a sensitive and efficient method for screening BACE1 inhibitors with the robustness of HTRF technology. HTRF assays offer many advantages over other technologies: Homogeneous add-and-read format No wash steps Low background Straightforward miniaturization from 96- or 384-well microplates to high density assay formats such as 384-well low volume and 1536-well plates Stable signal, providing flexibility in time of readout or size of assays
Assay principle The HTRF BACE1 assay is based on our amyloid precursor protein (APP) substrate, labeled with both biotin and Eu3+ Cryptate. Active BACE1 enzyme cleaves the substrate, causing the loss of energy transfer, and thus a reduction in SA-XL665 or Sa-d2 emissions. Assay protocol The assay is run in a two-step process: 1.BACE1 enzymatic reaction: Mix together the APP substrate, BACE1 and/or compounds, then incubate (we recommend a time course study to determine the optimal stimulation time). 2.BACE1 activity detection: Add Streptavidin-XL665 or Streptavidin-d2.