Sodium Butyrate Protocol
Application
Sodium butyrate (Millipore cat. no.TR-1008-G) can act to increase the efficiency of transfection and expression for both transient and stable transfections. It increases enhancer-dependant transcription of inserts driven by the SV40 early promoter. Vectors of this type include any of the pSV and pSV2 plasmids. Treatment of transfected cells with sodium butyrate can result in: a 2–3 fold increase in the number of cells able to express the insert; a 10–30 fold increase in the number of stable transfected colonies; and cells treated with sodium butyrate at the time of transfection can be re-introduced by a second treatment weeks after the initial treatment and yield a 4–7 fold increase in transcription of the SV40 early promoter driven insert.This treatment can be used on cells transfected by the Calcium Phosphate, DEAE Dextran and electroporation method. The stock solution is 500 mM sodium butyrate.
Note: After treatment with sodium butyrate the morphology of some cell types will be altered (for example CV-1 cells will become swollen and flattened) and the cell density will decrease by 10 to 50%. One needs to test each cell type for the best concentration of sodium butyrate to treat with; however, following is a list of some commonly used cell types and the final concentration of sodium butyrate.
| Cell Type | Final concentration of sodium butyrate |
| CV-1, COS-7 | 10mM |
| NIH-3T3 | 7mM |
| Hela | 5mM |
| CHO | 2mM |
Protocol
- Transfect cells by Calcium Phosphate or DEAE Dextran or electroporation.
- For cells transfected by Calcium Phosphate method; remove media with CaPO4-DNA, wash cells with medium without serum, add glycerol shock solution for 3 minutes at room temperature, wash cells with medium without serum, and add fresh medium with serum.
- For cells transfected by DEAE Dextran or electroporation; after treatment with DNA, feed cells fresh medium with serum.
- Add sodium butyrate to medium and incubate cells for 12 to 14 hours.
- Remove medium with sodium butyrate, wash cells with medium without serum, feed cells fresh medium with serum and incubate.
- Harvest cells for transient expression as desired. For stable expression, 24 hours after step #5, place cells into appropriate selection medium.
Reference
Gorman, et al., Nucleic Acids Research 1983; 11:21.
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