Expression of chemokine receptor in RBL-2H3 cells
ming88us at yahoo.com
Mon Sep 9 12:36:46 EST 2002
I'm working with some chemokine receptors and try to
establish stable RBL-2H3 cell lines expressing them.
The cDNAs were cloned into pcDNA3.1(ECoRI and XbaI)
and the recombinant plasmid (digested with Bgl II) was
transfected into RBL-2H3 cells by electroporation.
After 3 weeks of selection with 800ug/ml G418, cells
were pooled and stained with PE-conjugated monoclonal
antibody to detect the receptor expression on the cell
surface. Unfortunately we did not get any positive
cells on FACS, although DNA sequencing and RT-PCR
results were fine. BTW, the antibody was good, since
it worked very well with human monocytes.
What shall I do? Any suggestion is welcome.
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