Product Qualification

Lip2000™ has been extensively tested by transfection of HEK293 cells with an EGFP reporter containing plasmid. Lip2000™ is free of microbial contamination.

Important Guidelines

Follow these guidelines when performing transfections:

1. The ratio of DNA (in µg) : Lip2000™ (in µl) to use when preparing complexes should be

1:2 to 1:3 for most cell lines. To transfect 0.5 -2 X105 cells in a 24-well format, use 0.8-1 µg DNA

and 2-3 µl of Lip2000™. Optimizing transfection by varying DNA/Lip2000™ ratio is possible.

2. It is CRITICAL to transfect cells at high cell density. 90-95% confluence the time of transfection is recommended to obtain high efficiency and expression levels and to minimize decreased cell growth associated with high transfection activity. Lower cell densities are suitable with optimization of conditions. Take care to maintain a standard seeding protocol between experiments because transfection efficiency is dependent on culture confluence.

3. DO NOT add antibiotics to media during transfection as this will cause cell death.

Optimizing Transfection

To obtain the highest transfection efficiency and low non-specific effects，optimize transfection conditions by varying DNA and Lip2000™ concentrations， and cell number. Make sure that cells are greater than 90% confluent and vary DNA (µg) : Lip2000™ (µl) ratios from 1:0.5 to 1:5