By Edouard Kurstak
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The standard units obtained are directly proportional to antibody activity. 52 4. , 1:1000) against their respective titers. The test samples in subsequent experiments are measured for activity at the same dilution (internal standards from the original curve can be included) and the titer is obtained directly from the standard curve. Malvano et al. (1982) used a similar approach, except that instead of a battery of different sera, one positive serum, calibrated against a WHO standard serum, diluted to varying degrees with a negative serum was used to construct the standard-unit curve.
After fixation it is necessary to wash the preparation in cold saline to attain neutrality. Numerous formulations of cross-linking fixatives have been described. In particular formaldehyde is popular, whereas glutaradlehyde (a dialdehyde) yields increased cross-linking, but tends also to decrease antigenicity. The common commercial formaldehyde solution (37% formalin) is not suitable, since it contains a number of stabilizers (Fair and Nakane, 1981).
25 volume of 2 M sodium carbonate. Alkaline phosphatase should be assayed in a buffer depending on the origin of the enzyme. 8 (adjusted with hydrochloric acid) is used. -nitrophenylphosphate is added to these buffers at 1 mg/ml. The production of nitrophenol is monitored at 405 nm. 1 volume of 10 mM cysteine. Alkaline phosphatase is also useful for histochemical detection of antigens on nitrocellulose blots. Particularly useful are the reagents described by Blake et al (1984) and Knecht and Dimond (1984).
Enzyme Immunodiagnosis by Edouard Kurstak