Electrophoretic technique for assaying the relative distribution

Details Electrophoretic technique for assaying the relative distribution Electrophoretic technique for assaying the relative distribution

1979-05-29

1981-01-20

Prescott, Arthur C.

Chemistry: electrical and wave energy

Processes and products

Electrostatic field or electrical discharge

204180S, 204299R, 424 12, 23230B, 435 26, G01N 3316, G01N 2726

Patent

active

042460840

ABSTRACT:
An electrophoretic technique for assaying the relative distribution of lactate dehydrogenase (LD) isoenzymes characterized in that the electrophoretic and/or substrate buffer employed therein comprises a first and second moiety wherein the first moiety is selected from a group consisting of alkali metal 5,5-diethylbarbiturate, ammonium 5,5-diethylbarbiturate, and mixtures thereof, and wherein the second moiety is selected from a group consisting of bicine (N,N-bis(2-hydroxyethyl)glycine), tricine (N-tris(hydroxymethyl)methylglycine), and glycylglycine. The buffer has a pH of from about 7 to about 9 at 25.degree. C.
Preferably, the buffer further comprises a solid organic water soluble acid and either 2-amino-2-methyl-1,3-propanediol (AMPD) or 2-amino-2-hydroxymethyl-1,3-propanediol (Tris) or mixtures thereof.
Buffers within the scope of the instant invention, when compared to prior art buffers, increase the relative percentage of LD.sub.5 and thereby improve the accuracy and reproducibility of the electrophoretic data.

REFERENCES:
patent: 2555487 (1951-06-01), Haugaard et al.
patent: 3384564 (1968-05-01), Ornstein et al.
patent: 3582490 (1971-06-01), Zemel
patent: 3692654 (1972-09-01), Svendsen
patent: 4013513 (1977-03-01), Lederer
patent: 4139440 (1979-02-01), Chrambach et al.
patent: 4147606 (1979-04-01), Golias
Buhl et al., "A Search for the Best Buffer . . . . . ", Clin. Chem., 22(11): 1872-1875 (1976).
Gueffroy, Ed., "A Guide for the Preparation and Use of Buffers in Biological System", Calbiochem, LaJolla, CA. (1975).

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