Alkaline phosphatase isoenzyme determination
AssigneeSigma Chemical Company
InventorWilliam H. Starkweather
Life of this patent
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Abstract
Alkaline phosphatase isoenzymes are separated electrophoretically in agarose gels, or cellulose acetate membranes, equilibrated in a solution containing a low ionic strength buffer and a non-ionic detergent, such as Triton X-100. The electrophoretically separated isoenzymes are developed colorimetrically using 5-bromo-4-chloro-3-indolyl phosphate with a transphosphorylating buffer. Zones of enzyme activity appear as sharply-defined brilliant blue bands. The method is capable of resolving liver, bone, placental, intestinal and bile isoenzymes.
Claims18
In an electrophoretic process of determining alkaline phosphatase isoenzymes in a sample comprising a step of applying said sample to a support medium, a step of subjecting the support medium to a differential electric potential to separate said isoenzymes, and a step of rendering said isoenzymes detectable, the improvement wherein said support medium is chosen from the group consisting of agarose gels and cellulose acetate membranes, said support medium containing a buffer and a non-ionic detergent diffused through it.
An electrophoretic method capable of separating into well-defined, easily observed, discrete fractions at least the liver, bone, placental, intestinal and bile alkaline phosphatase isoenzymes in a biological fluid, said method comprising a step of applying a sample of said fluid to a support medium containing a buffer and a non-ionic detergent;
- imposing a differential electric potential across the support medium;
- and a subsequent dyeing step, said dyeing step comprising the addition of a phosphate ester of an indigo congener stain, an activating amount of a soluble magnesium salt and a transphosphorylating buffer.
Dependencies are inferred from the claim text.