Micro ELISA for measurement of parathymosin alpha utilizing a monoclonal antibody.
A monoclonal antibody to the non-thymosin alpha 1 overlapping sequence of parathymosin alpha was used to develop a micro ELISA to measure parathymosin alpha. Parathymosin alpha levels were measurable in both mouse tissue extracts and in human adult and cord serum. To measure parathymosin alpha, a 40% ammonium sulfate precipitated monoclonal antibody specific for the parathymosin alpha fragment was preincubated with the parathymosin alpha fragment or samples for 4 hours at 4 degrees C, incubated an additional 24 hours in microtiter plates coated with the parathymosin alpha fragment and then assayed by the biotin-avidin-alkaline phosphatase method. Using the assay, parathymosin alpha could be measured over a range of 5 to 630 ng/ml and cross-reactivity with thymosin alpha 1 was not observed. The parathymosin alpha level was 400 +/- 140 (mean +/- SD) ng/ml in normal human sera (n = 12) and 150 +/- 80 ng/ml in cord sera (n = 12). The concentration of parathymosin alpha was highest in tissue extracts of mouse liver (153 +/- 40 micrograms parathymosin alpha per g of tissue), kidney (125 +/- 33), and lung (105 +/- 38). Levels were lowest in thymus (81 +/- 20), spleen (83 +/- 23), and brain (71 +/- 23).
Wada
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Department of Biochemistry and Molecular Biology, George Washington University School of Medicine and Health Sciences, Washington, D.C. 20037.
Naylor
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Naylor
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Goldstein
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D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis*; D001923:Brain Chemistry; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005312:Fetal Blood / Q000032:analysis; D006801:Humans; D066298:In Vitro Techniques; D007668:Kidney / Q000032:analysis; D008099:Liver / Q000032:analysis; D008168:Lung / Q000032:analysis; D051379:Mice; D013154:Spleen / Q000032:analysis; D013947:Thymosin / Q000031:analogs & derivatives* / Q000032:analysis / Q000097:blood; D013950:Thymus Gland / Q000032:analysis
D000911:Antibodies, Monoclonal; D013947:Thymosin; C044498:parathymosin alpha