Does anyone know which cytokine receptors are expressed by these cell lines? I would like to do PCR on them to check the expression of several receptors to get an idea if cytokine treatments do not have an effect because of lacking expression. But we do not have any primers available for this purpose. If I design new primers I would need some positive controls to validate them... Any suggestions for this issues?
Thanks!
Hi Navina.
Why not design the primer pairs you are interested in and validate them on murine splenocytes - either fresh or LPS stimulated?
One thing you may find useful is to design your primer pairs to have optimal efficiency under the same PCR conditions on cDNA. In this way, you will be able to mix and match your primers on the same plate, should you need to.
Good Luck
G
Helo Grant,
thank you very much for your answer.
Do you know if murine splenocytes express all of the following receptors.? Receptors for: IFNg, TNFa, IL4, IL6, IL10?
Thanks again,
Navina
You can use cytokine/cytokine receptor arrays with cell extracts.
R and D systems sells the arrays. Ray Biotech can do a 308 different protein array with cell lysates/extracts. The protein arrays contain all sorts of proteins including many cytokine receptors, such as TNFRI and II. IL-1RA, IFN receptors, etc, etc.
Thank you Grant! I will give it a try!
Marcia, I was looking into arrays but the one I found was really expensive. :(
The arrays are expensive. The Ray Biotech one costs about 1000 dollars, maybe for two samples. But the R and D one is better..
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