Method for extracting quantitative information relating to interactions between cellular components
US7282347B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Jul 3, 2001 |
| Grant date | Oct 16, 2007 |
| Priority date | — |
| Expiry date | May 26, 2022 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N33/68
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
A method is described to assay for protein interactions in living cells, e.g. by the introduction of two heterologous conjugates into the cell. The method uses the measurement of cellular distribution of a detectable component (e.g. a GFP-labeled˜fluorescent probe) to indicate the presence or absence of an interaction between that component and a second component of interest. The method uses the knowledge that certain components can be stimulated to redistribute within the cell to defined locations. Inducible redistribution systems make it possible to determine if specific interactions occur between components. Inducible systems are described where it is demonstrated that the redistribution stimuli are essentially “null”, in that they affect no other system in the cell during the assay period, other than the component whose redistribution can be induced. Also described is an extraction buffer which is useful in high throughput screening for drugs which affect the intracellular distribution of intracellular components. The extraction buffer comprises a cellular fixation agent and cellular permeabilisation agent. Optimizing the composition of the extraction buffer and its application…
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.