How do I enhance surface expression of FPR1 on primary human neutrophils?

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Mahendra Chordia

You may able to prime the cells with fMLP or LPS or TNF-alfa. If you are just going to test antibody for in vitro titration experiment incubate about 0.1 to 10 nanomolar fMLP for FPR receptor expression on isolated neutrophils for about 1h. Measure antibody binding with and without fMLP stimulation.

Landon W Locke

Thanks Mahendra. After reading some older papers, I think I will choose 50ng/mL PMA for neutrophil stimulation for 5 min at 37C. PMA seems to increase surface expression of the FPR.

Yes PMA should work fine too. Li Xiao reported ear lobe inflammation model for imaging using PMA, check the molecular imaging paper.

Elisabeth Weiß

There are some puplications by Ji Ming Wang about the upregulation of FPR receptors in microglia cells. I used Pam2Cys to induce FPR1 and FPR2 expression in mouse neutrophils. Which antibody do you use? I´m searching for a good one.