摘要:
Methods and compositions are provided for detecting molecular translocations, particularly protein translocations within and between subcellular copartments, using at least two components that exhibit a localization-dependent difference in complementation activity. In particular, alpha-complementing β-galactosidase fragments are provided. These β-galactosidase reporter fragments display significantly enhanced enzymatic activity when one fragment is localized in a membrane. Methods for carrying out no-wash ELISA assays based on the reporter component system are also provided.
摘要:
Methods and compositions for detecting molecular interactions are provided. Aspects of the invention include the use of a reduced affinity enzyme complementation reporter system. Also provided are systems and kits for use in practicing embodiments of the methods.
摘要:
Methods and compositions for producing hRNA, e.g., shRNA, expression modules for specific target nucleic acids are provided. In the subject methods, an initial nucleic acid, e.g., dsDNA, synthetic DNA, etc., corresponding to the target nucleic acid of interest is converted to an intermediate nucleic acid. The resultant intermediate nucleic acid, following an optional size modification step, is then converted to a linear dsDNA that includes at least one copy of the hRNA expression module of interest, or a precursor (i.e., pro-shRNA expression module) thereof, where in certain embodiments conversion may include amplification. Also provided are reagents, systems and kits for use in practicing the subject methods. The subject methods and compositions find use in a variety of different applications.
摘要:
Sequential reporter enzyme luminescence (SRL) methods are provided. In the subject methods, the activity of a reporter enzyme is evaluated using a secondary reporter system that employs a product of a reporter enzyme mediated reaction as a luminescent substrate, e.g., luciferase substrate. Also provided are kits and other compositions that find use in practicing the subject methods.
摘要:
The disclosure provides, inter alia, methods for enhancing the contribution of circulating stem cells to a target tissue. Such methods may be useful for treating a variety of disorders. In a preferred embodiments, circulating stem cell contribution is enhanced by causing damage to the target tissue, or by administering an agent that mimics an aspect of a damage response. The disclosure also provides methods for monitoring the contribution of circulating stem cells to a target tissue, and for developing agents that modulate such contribution.