Detalles de config. de Preguntas frecuentes
-
Can I detect the direct binding of a ligand to a cell? What do I measure in a cell-based assay?
-
In cell based assays Epic technology does not measure the direct binding of ligand to receptor. The technology measures the downstream, integrated physiological response of the cell to the binding/activation of the biological target within a sensing region of ~150 nm from the sensor surface. We refer to the movement of intracellular components in response to a cellular stimulus, the Dynamic Mass Redistribution (DMR). Responses are measured as wavelength shifts (units of pm) in the reflected light from the bottom of the optical biosensor.
Detalles de config. de Preguntas frecuentes
-
What is DMR?
-
DMR stands for Dynamic Mass Redistribution and refers to the movement of intracellular components in response to a cellular stimulus.
Detalles de config. de Preguntas frecuentes
-
Can I test multiple cell lines on a single plate?
-
Yes, as long as the cell culture conditions are similar. We have successfully cultured multiple cell lines in adjacent columns on a single plate and tested their response to various stimuli.
Detalles de config. de Preguntas frecuentes
-
What are the important parameters to optimize in a cell-based assay?
-
The most important parameters to optimize for each new cell assay are: 1) cell seeding density; 2) microplate type (uncoated, fibronectin-coated, or self-coated); 3) culture conditions; 4) liquid handling parameters for compound dispensing and mixing.
Detalles de config. de Preguntas frecuentes
-
How do I choose the correct plate for a cell-based assay?
-
For loosely-adherent cells, we recommend fibronectin-coated microplates. For strongly adherent cells and suspension cells, we recommend that you evaluate both fibronectin-coated and uncoated microplates.
Detalles de config. de Preguntas frecuentes
-
Can Epic Technology be used to study suspension cells?
-
Yes. Epic Technology has successfully been used to study both adherent and non-adherent (suspension) cells. Liquid handling parameters for compound addition may require optimization when working with suspension cells.
Detalles de config. de Preguntas frecuentes
-
Are Epic cell-based assays sensitive to cell density?
-
Yes. In general we recommend running assays with a confluent monolayer. For both adherent and non-adherent cells, the optimal seeding density should be empirically determined during assay optimization.
Detalles de config. de Preguntas frecuentes
-
Does the Epic signal depend upon the expression level of the receptor?
-
Yes. However, for a given cell type, there is a maximal signal one can obtain for a particular receptor. The expression threshold required to reach the maximal signal is dependent on (1) the receptor; (2) the cell type; (3) the ligand; (4) the signaling capacity of the receptor; and (5) the interaction of the receptor with other proteins.