The development of a radioimmunoassay procedure for the estimation of Tamm-Horsfall glycoprotein in human serum.
1. A quantitative radioimmunoassay was developed for the measurement of nanogram amounts of Tamm-Horsfall (TH) glycoprotein in the presence of serum proteins at concentrations above 30 mg/ml. 2. Specific anti-(TH-glycoprotein) antibodies were labelled with 125I and these were usable for a period of 8 weeks. 3. Agarose beads (Sepharose 4B), to which TH-glycoprotein had been coupled via cyanogen bromide activation of the Sepharose, were used as the solid phase in the assay. This proved stable upon storage at 4 degrees C for periods in excess of 4 months. 4. The dissociation of the glycoprotein in the presence of serum proteins that was necessary for quantification was achieved by subjecting the sample to ultrasonication. 5. Assays conducted on a small sample of normal serum produced evidence that normal serum contained amounts (50-180 ng/ml) of a substance that reacted similarly to TH-glycoprotein in the assay procedure and in a series of experiments conducted to confirm the presence of this substance in human serum.
D000906:Antibodies / Q000302:isolation & purification; D001798:Blood Proteins / Q000276:immunology; D006023:Glycoproteins / Q000097:blood* / Q000276:immunology; D006801:Humans; D008297:Male; D011863:Radioimmunoassay / Q000379:methods*; D011864:Radioimmunosorbent Test / Q000379:methods*
D000906:Antibodies; D001798:Blood Proteins; D006023:Glycoproteins