Jul 25, 2023

Public workspaceHEK293 cell culture for co-immunoprecipitation experiments

DOI
dx.doi.org/10.17504/protocols.io.kxygx3zeog8j/v1
  • 1University of Pennsylvania
Dan Dou
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DOI: dx.doi.org/10.17504/protocols.io.kxygx3zeog8j/v1
Protocol Citation: Dan Dou, Erika Holzbaur 2023. HEK293 cell culture for co-immunoprecipitation experiments. protocols.io https://dx.doi.org/10.17504/protocols.io.kxygx3zeog8j/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: July 20, 2023
Last Modified: May 31, 2024
Protocol Integer ID: 85308
Keywords: ASAPCRN
Funders Acknowledgement:
ASAP
Grant ID: ASAP-000350
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)
Culture HEK293 cells in DMEM (Corning) supplemented with 10% fetal bovine serum (HyClone)
Maintain cells at 37 degrees Celsius with 5% CO2
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
24 hours later, transfect using FuGENE 6 (6-12 µg total plasmid DNA; Promega)