Development of high-throughput radioligand binding assays for interleukin-1 alpha (IL-1 alpha) and tumor necrosis factor (TNF-alpha) in isolated membrane preparations.
Cytokine binding has been studied in a variety of intact cells, and in isolated receptor preparations. Each approach is associated with limitations with regard to screening large numbers of samples on a repetitive basis. In order to provide a more reproducible system of screening for compounds which modify IL-1 alpha and TNF-alpha binding, we have developed isolated membrane preparations for studying agents which can alter the association of these ligands with their receptors. These results demonstrate IL-1 alpha binding to BALB/c 3T3 cell membranes and TNF-alpha binding to HeLa S3 cell membranes, and indicate that this is a viable approach to high-throughput screening.
Maloff
B L
BL
Pharmacology Services, Panlabs, Inc., Bothell, WA 98011.
Delmendo
R E
RE
D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry; D007375:Interleukin-1 / Q000032:analysis*; D007457:Iodine Radioisotopes; D007700:Kinetics; D011869:Radioligand Assay / Q000379:methods*; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis*
D007375:Interleukin-1; D007457:Iodine Radioisotopes; D014409:Tumor Necrosis Factor-alpha