We have difficulties with a neuropharmacological assay.
We use HEK-293 cells stably transfected with the human receptor TRPV1. We monitor intracellular calcium using Fura2.
We stimulate these cells (coated on 96-well plate) with capsaicin (10-100 µM) and measure the fluorescence with a FLEX station. We are unable to observe an increase of intra Ca2+.
Does anyone have any advice on possible experimental precautions to be taken in this assay?
Thank you very much.