384 Well Microplate Analysis using the BacMam Histone H3 [pSer10] Cellular Assay Kit in the Corning® HYPERFlask® Cell Culture Vessel
Pardo, A.M P.1, Gainer, T.G.2 and Rothenberg, M.E.1
1Corning Incorporated, Life Sciences 2 Alfred Road, Kennebunk, ME, 2Life Technologies, 501 Charmany Drive, Madison, WI
Abstract:The high throughput analysis of potential drug candidates in a cell-based assay screen is, to say the least, complicated. New technologies are currently on the market to aid the assay biologist in developing and screening their drug candidates. Two such technologies include the BacMam Histone H3 [pSer10] Cellular Assay Kit (Life Technologies Cat. #A12898) and Corning's High Yield PERformance Flask (HYPERFlask) cell culture vessel. The baculovirus-mediated gene delivery system or BacMam in combination with the LanthaScreen® technology can be used for the analysis of specific posttranslational modifications of histones. The HYPERFlask vessel is a multilayer flask that uses a gas permeable film to provide efficient gas exchange between the cells, culture medium and the environment surrounding it. The experiments presented in this poster describe a protocol combining both technologies using HEK293 cells as the model. HEK293 cells containing the GFP-Histone H3 fusion gene were seeded into Corning white 384 well normal (Corning Cat.#3570) and low volume (Corning Cat.#3826) flat bottom microplates and assayed for LanthaScreen® activity using the PerkinElmer® Envision® plate reader containing Life Technologies supplied Terbium filters. The data show successful transduction of the HEK293 cells with the GFP-Histone H3 fusion gene utilizing both technologies and resulting in a high yield production of functional GFP-Histone H3 transduced cells.
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