Hello everybody,
I'm doing a ChIP on an HA-tagged NFkB tranfected in NIH 3T3 cells.
For the qPCR I need a gene that is expressed in this cells and whose promoter is bound by NFkB. This will be my positive control.
Thank you
Hi,
My first suggestion would be to check the company you get your antibody from and see if they provide positive control primers. If they do not, I would also search for papers that have used the same antibody for ChIP. Because NFkB bind to a number of genes, it should be quick to find a suitable promoter, mainly in inflammatory related genes. A good example would be TNFa or IkBa.
Tnip1 seems like a good gene for you. There are supposed to be 5 NFkB binding sites. You should be able to design your own primers starting with Chromosome RefSeq #: NC_000077.5 where the transcriptional start site is at position: 54776414. Then there are supposed to be NFkB binding sites at positions 54770307, 54771923, 54774705, 54775690, and 54785701. A program like Primer-Blast might be useful.
Thank you!!
Do you have in any case the literature reference for these binding sites?
- Badges
- Science topic
- Similar topics
- Biological Science
- Genetics
- Gene Expression