A radioreceptor assay for mass measurement of inositol (1,4,5)-trisphosphate using saponin-permeabilized outdated human platelets. 1998

J Rivera, and M L Lozano, and R Gonzalez-Conejero, and J Corral, and F De Arriba, and V Vicente
Unit of Hematology and Hemotherapy, School of Medicine, Hospital General Universitario, Murcia, Spain.

The binding of inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] to saponin-permeabilized blood-bank-outdated human platelets, 6 days old, has been characterized (Kd = 3.8 nM; Bmax = 1.7 pmol/mg protein) and used to develop a novel radioreceptor assay which allows the measurement of the Ins(1,4,5)P3 content in resting or agonist-stimulated cells. This assay is as sensitive (0.25 pmol in a 0.25 ml volume), specific, and reproducible as previously proposed methods. In addition, obtaining large batches of the Ins(1,4,5)P3 binding protein by treating outdated platelets with saponin is simple and quick and uses otherwise discarded material. Moreover, the assay is considerably cheaper than commercially available kits. Using this method we confirmed that thrombin evokes a rapid, transient, and dose-dependent increase in the platelet concentration of Ins(1,4,5)P3.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D010539 Permeability Property of membranes and other structures to permit passage of light, heat, gases, liquids, metabolites, and mineral ions. Permeabilities
D011869 Radioligand Assay Quantitative determination of receptor (binding) proteins in body fluids or tissue using radioactively labeled binding reagents (e.g., antibodies, intracellular receptors, plasma binders). Protein-Binding Radioassay,Radioreceptor Assay,Assay, Radioligand,Assay, Radioreceptor,Assays, Radioligand,Assays, Radioreceptor,Protein Binding Radioassay,Protein-Binding Radioassays,Radioassay, Protein-Binding,Radioassays, Protein-Binding,Radioligand Assays,Radioreceptor Assays
D001792 Blood Platelets Non-nucleated disk-shaped cells formed in the megakaryocyte and found in the blood of all mammals. They are mainly involved in blood coagulation. Platelets,Thrombocytes,Blood Platelet,Platelet,Platelet, Blood,Platelets, Blood,Thrombocyte
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D012503 Saponins A type of glycoside widely distributed in plants. Each consists of a sapogenin as the aglycone moiety, and a sugar. The sapogenin may be a steroid or a triterpene and the sugar may be glucose, galactose, a pentose, or a methylpentose. Saponin
D012680 Sensitivity and Specificity Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed) Specificity,Sensitivity,Specificity and Sensitivity
D013917 Thrombin An enzyme formed from PROTHROMBIN that converts FIBRINOGEN to FIBRIN. Thrombase,Thrombin JMI,Thrombin-JMI,Thrombinar,Thrombostat,alpha-Thrombin,beta,gamma-Thrombin,beta-Thrombin,gamma-Thrombin,JMI, Thrombin
D015203 Reproducibility of Results The statistical reproducibility of measurements (often in a clinical context), including the testing of instrumentation or techniques to obtain reproducible results. The concept includes reproducibility of physiological measurements, which may be used to develop rules to assess probability or prognosis, or response to a stimulus; reproducibility of occurrence of a condition; and reproducibility of experimental results. Reliability and Validity,Reliability of Result,Reproducibility Of Result,Reproducibility of Finding,Validity of Result,Validity of Results,Face Validity,Reliability (Epidemiology),Reliability of Results,Reproducibility of Findings,Test-Retest Reliability,Validity (Epidemiology),Finding Reproducibilities,Finding Reproducibility,Of Result, Reproducibility,Of Results, Reproducibility,Reliabilities, Test-Retest,Reliability, Test-Retest,Result Reliabilities,Result Reliability,Result Validities,Result Validity,Result, Reproducibility Of,Results, Reproducibility Of,Test Retest Reliability,Validity and Reliability,Validity, Face
D015544 Inositol 1,4,5-Trisphosphate Intracellular messenger formed by the action of phospholipase C on phosphatidylinositol 4,5-bisphosphate, which is one of the phospholipids that make up the cell membrane. Inositol 1,4,5-trisphosphate is released into the cytoplasm where it releases calcium ions from internal stores within the cell's endoplasmic reticulum. These calcium ions stimulate the activity of B kinase or calmodulin. 1,4,5-InsP3,Inositol 1,4,5-Triphosphate,Myo-Inositol 1,4,5-Trisphosphate,1,4,5-IP3,Myoinositol 1,4,5-Triphosphate

Related Publications

J Rivera, and M L Lozano, and R Gonzalez-Conejero, and J Corral, and F De Arriba, and V Vicente
January 2013, Methods in molecular biology (Clifton, N.J.),
J Rivera, and M L Lozano, and R Gonzalez-Conejero, and J Corral, and F De Arriba, and V Vicente
January 2006, Methods in molecular biology (Clifton, N.J.),
J Rivera, and M L Lozano, and R Gonzalez-Conejero, and J Corral, and F De Arriba, and V Vicente
January 1999, Methods in molecular biology (Clifton, N.J.),
J Rivera, and M L Lozano, and R Gonzalez-Conejero, and J Corral, and F De Arriba, and V Vicente
January 2005, Methods in molecular biology (Clifton, N.J.),
J Rivera, and M L Lozano, and R Gonzalez-Conejero, and J Corral, and F De Arriba, and V Vicente
December 1984, Proceedings of the National Academy of Sciences of the United States of America,
J Rivera, and M L Lozano, and R Gonzalez-Conejero, and J Corral, and F De Arriba, and V Vicente
March 1992, The Journal of pharmacology and experimental therapeutics,
J Rivera, and M L Lozano, and R Gonzalez-Conejero, and J Corral, and F De Arriba, and V Vicente
February 1986, The Biochemical journal,
J Rivera, and M L Lozano, and R Gonzalez-Conejero, and J Corral, and F De Arriba, and V Vicente
April 1986, The Journal of biological chemistry,
J Rivera, and M L Lozano, and R Gonzalez-Conejero, and J Corral, and F De Arriba, and V Vicente
November 1985, Thrombosis research,
J Rivera, and M L Lozano, and R Gonzalez-Conejero, and J Corral, and F De Arriba, and V Vicente
January 1998, Methods in molecular biology (Clifton, N.J.),
Copied contents to your clipboard!