I am trying to find reference genes that can be used, the cells are being stimulated with LPS,however the reference genes I have used so far (beta 2 microglobulin and beta actin) have been shown to be highly unstable. Does any one have any recommendations?
I found this paper which suggests GADPH is unstable http://www.ncbi.nlm.nih.gov/pubmed/15182252. Ill have a look into Ppia thanks
We have used mouse actin and observed no differences in expression levels with qRT-PCR with LPS stimulation for upto 24h.
I use HPRT, mainly because it is express at lower levels than most other house keeping genes. It is very stable in my in vivo models. I have not tested its stability after a strong in vitro LPS stimulation.
The attached publication might be helpful.
In my previous lab, we used 18S mRNA for macrophages stimulated with LPS.
The following publication may be helpful:
Autophagy regulates phagocytosis by modulating the expression of scavenger receptors.
Bonilla DL et al
Immunity. 2013 Sep 19;39(3):537-47
PMID: 24035364
(The information is provided in the Experimental Procedures section and in Supplementary figure S5)
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