TY - JOUR
T1 - [7] Assay of Cyclic Nucleotides by Receptor Protein Binding Displacement
AU - Gilman, Alfred G.
AU - Murad, Ferid
N1 - Funding Information:
These studies were supported in part by United States Public Health Service grants NS10193 and AM15316 and the Virginia Heart Association. Ferid Murad is the recipient of USPHS Research Career Development Award AM70456.
PY - 1974/1/1
Y1 - 1974/1/1
N2 - Receptor protein binding displacement assays for cyclic nucleotides are based on competition for protein binding sites between radioisotopically labeled nucleotide and the unlabeled material to be quantified. The binding proteins utilized are those that occur naturally and that interact with the appropriate cyclic nueleotide with high affinity. These procedures offer intrinsic advantages of simplicity of operation, high sensitivity, and high specificity. These features are particularly marked in the case of the assay for adenosine 3', 5'-cyclic monophosphate (cyclic AMP, cAMP) and have led to considerable utilization of this method. Binding activity may be simply followed through the purification procedure by incubation of small aliquots (5–25μl) of appropriate fractions with an excess of [3H] cAMP in 20 mM potassium phosphate, pH 6–7, for approximately 5 minutes at 30–37°C.
AB - Receptor protein binding displacement assays for cyclic nucleotides are based on competition for protein binding sites between radioisotopically labeled nucleotide and the unlabeled material to be quantified. The binding proteins utilized are those that occur naturally and that interact with the appropriate cyclic nueleotide with high affinity. These procedures offer intrinsic advantages of simplicity of operation, high sensitivity, and high specificity. These features are particularly marked in the case of the assay for adenosine 3', 5'-cyclic monophosphate (cyclic AMP, cAMP) and have led to considerable utilization of this method. Binding activity may be simply followed through the purification procedure by incubation of small aliquots (5–25μl) of appropriate fractions with an excess of [3H] cAMP in 20 mM potassium phosphate, pH 6–7, for approximately 5 minutes at 30–37°C.
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U2 - 10.1016/0076-6879(74)38010-X
DO - 10.1016/0076-6879(74)38010-X
M3 - Article
C2 - 4375777
AN - SCOPUS:0016223673
SN - 0076-6879
VL - 38
SP - 49
EP - 61
JO - Methods in Enzymology
JF - Methods in Enzymology
IS - C
ER -