HEK293 cell culture for co-immunoprecipitation experiments
Erika Holzbaur, Dan Dou
Published: 2023-07-25 DOI: 10.17504/protocols.io.kxygx3zeog8j/v1
Abstract
Here, we describe HEK293 cell culture for the purpose of co-immunoprecipitation experiments
Before start
Cells should be routinely tested for mycoplasma contamination using MycoAlert detection kit (Lonza, LT07)
Steps
1.
Culture HEK293 cells in DMEM (Corning) supplemented with 10% fetal bovine serum (HyClone)
2.
Maintain cells at 37 degrees Celsius with 5% CO2
3.
In preparation for co-immunoprecipitation experiments, passage HEK cells with Trypsin onto three 10 cm tissue culture dishes per experimental condition, at a density of 1:10
4.
24 hours later, transfect using FuGENE 6 (6-12 µg total plasmid DNA; Promega)
