I have a list of primers for human corneal keratocyte and endothelial cells. I want to validate these primers using cells that are known to express my genes of interest. Any suggestions on how I should approach this in terms of looking for cell lines to validate these primers.
Also, would the cell line have to human or can it be another specie?
Any insight would be much appreciated.
My recommendation would be to check here:
http://www.uniprot.org/
If you enter your gene of interest on top of page and you look at the expression after you pressed the entry number (pick the one for humans), they usually tell you in which kind of tissue it is expressed the most.
This gives you a hint. Otherwise look at antibody datasheets to see which cell line they have used to show that it is specific. If the protein is there the mRNA should be there as well.
I would choose human cell lines as well, since primers are mostly not 100% homologous between species.
Good luck
Lines Névine Eltonsy, , Vivian Gabisi, Xuesong Li K. Blair Russe Gordon B. Mills and Katherine Stemke-Hale. Detection Algorithm for the Validation of Human Cell . Int J Cancer. 2012 September 15; 131(6): E1024–E1030. doi:10.1002/ijc.27533.
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