IP3 accumulation in macrophages was measured using the HitHunter IP3 Fluorescence Polarization Assay (GE Healthcare, 90-0037-02). M0 macrophages were cultured in RPMI 1640 containing 10% FBS and M-CSF (100 ng/ml) and serum-starved overnight, and a series of different concentrations of potential endogenous agonists were added to designated wells in a 96-well plate. The reaction was quenched after 1 min, and the tracer was added, followed by an IP3-binding protein. Fluorescence was excited with a wavelength of 483 nm, and emission was measured at 530 nm.

Note: The content above has been extracted from a research article, so it may not display correctly.



Q&A
Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.



We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.