Immunoquantitation of human placental alkaline phosphatase using radial immunodiffusion

✍ Scribed by George J. Doellgast

• Publisher: Elsevier Science
• Year: 1977
• Tongue: English
• Weight: 994 KB
• Volume: 82
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(77)90164-6

✦ Synopsis

A simple procedure is described for immunoquantitation of human placental alkaline phosphatase by radial immunodiffusion. Agarose gels in petri dishes were overlayed with diluted antiserum, and, after equilibration of the antibody with the gel, the overlay was poured off and the gel was used for quantitation of enzyme protein using a specific stain to amplify visualization. The method has a variation of O-8.5% for triplicate determinations on a single plate, with a mean of 2.6%. Variations between plates were O-13%, with a mean of 4.2%. By including Triton X-100 detergent in the overlay solution, it was possible to quantitate the amount of enzyme in membrane preparations from placentae and cancer fluids. A lOOO-fold range of antiserum and enzyme dilutions was tested; over this range, only a 2-3 x deviation from the expected ring size was found. As little as 5 ng of enzyme protein could be measured by this technique.