I work as a tech (in training at the moment) at a research lab looking at the CRTh2 gene and it's role in asthma and other inflammatory diseases. At the moment we are running into a few problems in our efforts to optimize some experiments. Please take a look and contribute any advice or thoughts. If you need more info/details please just ask as I will attempt to be brief, and am no expert. I am excited to see what online discussion can bring to the research table!
For now I'm wondering if anyone has any experience working with the IL-4 cytokine. We are having trouble detecting IL-4 in PGD2 treated CD4+ CRTh2+ cell supernatants using ELISAs. At the moment we are using Diaclone ELISA IL-4 Eli-pair Kits and getting very low detections even though standards go down to 1.1pg/ml. Does anyone know any good protocols/kits for IL-4 that have worked well? Any that have accurate detection at very low concentrations? We are trying different dilutions, new serum (we were using FBS and are testing if Human Serum makes a difference). Supernatants are from 2million cells/ml cultures, we are trying 4million cells/ml culture to see if IL-4 production gets bumped well into detectable ranges for our kits.
Please share your thoughts!