G - Physics – 01 – N
Patent
G - Physics
01
N
150/15.5
G01N 21/01 (2006.01) G01N 33/48 (2006.01)
Patent
CA 1072428
Abstract of the Disclosure A substrate medium subsequent to its removal from a sup- post means containing a separated isoenzyme is contacted with a reagent selected from a group consisting of a composition compris- ing from 90 to 100 weight percent alcohol per unit volume, said alcohol containing from one to six carbon atoms, and from zero to 10 weight percent urea per unit volume; ketones containing from three to eight carbon atoms; an inorganic salt solution comprising from about 90 to about 99.99 percent of a solvent selected from a group consisting of water and inert polar organic solvents and from about 0.01 to about 10 percent of an organic salt having a formula R(Y)2 wherein R is selected from a group consisting of Pb+2, Ca+2, Sr+2 and Ba+2 and wherein Y is selected from a group consisting of Cl-, N03-, and C104-, and mixtures thereof, so that the fluorescent product present in the substrate medium is preci- pitated in situ, thereby localizing its presence and increasing the fluorometric detection technique's sensitivity. When creatine phosphokinase enzyme is electrophoreti- cally separated into its isoenzyme constituents, the above fluor- ometric detection technique is further improved by employing an electrophoretic buffer comprising an acid having a formula HOOC(CH2)nCH(NH2)COOH wherein n is an integer from 1 to 4, tris- (hydroxymethyl)aminomethane, and water. This buffer has a pH of 7 to 8.5 at 23°C. and an ionic strength of 0.02 to 1.5.
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