Application
Calcium Phosphate Transfection Kit has been used:to enable transfectionto transfect Hek293T cellsto transfect H29D cells
Suitable for transient and stable transfection of DNA into cultured mammalian cells. The following cells have successfully been transfected using the calcium phosphate method:BAECBowels melanoma cellsCHO K1COS-7Fibroblasts (human embryonic, neo derm)HEK293Huh 7IMR-90LLC (Lewis Lung Carcinoma)NIH3T3PC-12PCI-13SH-Sy5YSK-Hep-1T47D
Components
The Calcium Phosphate Transfection Kit contains:5 ml 2.5M CaCl2 (C2052)25 ml 2x HEPES Buffered Saline (H1012)25 ml molecular biology grade water (W4502)
Features and Benefits
Suitable for transient and stable transfection Reproducible for a wide range of cell types Widely referenced Inexpensive
General description
Calcium phosphate transfection is a commonly used method for the introduction of DNA into eukaryotic cells. This technique has been used to obtain both transient and stable transfections in a wide variety of cell types.
Other Notes
The protocol was developed for transient transfection of CHO cells using pSV40-CAT plasmid as a reporter gene.
Principle
The procedure is based on slow mixing of HEPES-buffered saline containing sodium phosphate with a CaCl2 solution containing the DNA. A DNA-calcium phosphate co-precipitate forms, which adheres to the cell surface and is taken up by the cell, presumably by endocytosis.