A micro-radioimmunoassay for the measurement of intracellular cAMP.
We have developed a rapid and sensitive micro-radioimmunoassay for the detection of cellular cAMP. The assay employs a simple method for preparing cellular lysates that requires as few as 500 cells per data point. Cellular lysates are incubated with cAMP-specific antibody complexes and [125I]cAMP in 96-well plates, harvested onto glass fiber filters and analyzed using a commercial autoradiograph imaging system. This method provides significant savings in reagents and may be particularly useful in situations where only a limited number of cells are available.
Berg
W
W
Department of Medicine, University of Maryland Medical School, Baltimore 21201.
Hornbeck
P V
PV
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D002772:Cholera Toxin / Q000494:pharmacology; D000242:Cyclic AMP / Q000032:analysis*; D006801:Humans; D016393:Lymphoma, B-Cell / Q000737:chemistry; D008832:Microchemistry; D011863:Radioimmunoassay / Q000379:methods*; D014407:Tumor Cells, Cultured
D002772:Cholera Toxin; D000242:Cyclic AMP; D015056:1-Methyl-3-isobutylxanthine