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The Z-factor defines a characteristic parameter of the capability of hit identification for each given assay. The following categorization of HTS assay quality by the value of the Z-Factor is a modification of Table 1 shown in Zhang et al. (1999); [2] note that the Z-factor cannot exceed one.
Signal-to-background ratio, signal-to-noise ratio, signal window, assay variability ratio, and Z-factor have been adopted to evaluate data quality. [9] [12] Strictly standardized mean difference has recently been proposed for assessing data quality in HTS assays. [13] [14]
Rotating cell‑based ligand binding assay using radioactivity or fluorescence, is a recent method that measures molecular interactions in living cells in real-time. This method allows the characterization of the binding mechanism, as well as K d, k on and k off. This principle is being applied in several studies, mainly with protein ligands ...
A mobility shift assay is electrophoretic separation of a protein–DNA or protein–RNA mixture on a polyacrylamide or agarose gel for a short period (about 1.5-2 hr for a 15- to 20-cm gel). [4] The speed at which different molecules (and combinations thereof) move through the gel is determined by their size and charge, and to a lesser extent ...
This means that even though the transcription factor is split into two fragments, it can still activate transcription when the two fragments are indirectly connected. The most common screening approach is the yeast two-hybrid assay. In this approach the researcher knows where each prey is located on the used medium (agar plates).
A protocol for the MAT test, using cultured cells, is described in the European Pharmacopoeia. [16] A recent study employing genetically engineered monocytes was able to significantly enhance the sensitivity of monocyte-based detection assays by bringing down the assay-completion time from more than 20 hours to 2–3 hours. [17]
Reporter genes can be used to assay for the activity of a particular promoter in a cell or organism. [23] In this case there is no separate "gene of interest"; the reporter gene is simply placed under the control of the target promoter and the reporter gene product's activity is quantitatively measured.
Chromatin Immunoprecipitation sequencing, also known as ChIP-seq, is an experimental technique used to identify transcription factor binding events throughout an entire genome. Knowing how the proteins in the human body interact with DNA to regulate gene expression is a key component of our knowledge of human diseases and biological processes.