Center of Excellence in Nutrigenomics

Processing of Affymetrix slides (includes hybridization, washing, scanning)

Measure caspase 3/7 using a luminescent assay.

The process of removing excess cholesterol from the periphery for transport and removal at the liver (reverse cholesterol transport, RCT) is important in preventing atherosclerosis. The first step is cholesterol efflux from macrophages. In these assays we can examine the ability of drugs and compounds to affect this process or alternatively measure the ability of serum samples to provide carriers proteins. We have developed a non-radioactive methods that is amenable to high-throughput screening ( Zhang J., S. Cai, B. R. Peterson, P. M. Kris-Etherton, and J. P. V. Heuvel. 2011. Development of a cell-based, high-throughput screening assay for cholesterol efflux using a fluorescent mimic of cholesterol. Assay Drug Dev Technol 9:136-146.)

Mammalian cells will be transiently transfected with the necessary plasmids for performing nuclear receptor reporter assays. The cells will be frozen using Indigo's proprietary Cryomite technology. Quality control will be performed and provided to the client prior to shipping. Detailed instructions for the thawing and treatment of the assay will be provided. Available assays can be found at indigobiosciences.com

DNA is isolated, quantified and diluted

Cell culture setup. Includes cell line propagation, transfection and treatment. Experiments are performed in 96-well plates with a maximum of 10-plates per day. We can grow many different transformed cell lines, please inquire about availability.

Analysis, consulting and data interpretation

DNA is isolated, quantified and diluted

RNA extraction appropriate for real-time PCR

Real time PCR will be performed using SYBR green as the indicator in an ABI 7300 detection system.