Radioimmunoassay of rat submandibular salivary gland mucin

Abstract

A solid phase radioimmunoassay (RIA) for rat submandibular mucin (RSM) was developed and applied to studies of mucin concentration, antigenicity and secretion. The assay utilizes the affinity of 125I-Protein A for IgG to quantitate antigendashantibody immunocomplexes immobilized in the wells of plastic microtitre plates. The technique was highly reproducible and capable of detecting as little as 3 ng mucin protein. The submandibular glands of rats weighing 150–180 g contained 257 μg of mucin (dry wt) per mg protein, which is equivalent to about 3.6 mg of mucin per gland. The antiserum to RSM was cross-reactive with mucins from rat sublingual and pig submandibular glands, and rat, human and pig small intestine. No cross-reactivity was detected with mucins from mouse, canine or bovine submandibular glands, and there was no evidence that ABH blood-group sugars contributed to mucin antigenicity. The RIA was used to estimate secretion from dispersed rat submandibular gland cells and gave a more specific and accurate assay of mucin release than previous assays of precursor-labelled radioactive glycoproteins in the culture medium. The β-adrenergic agonist, isoproterenol, stimulated immunoreactive mucin secretion from cultured cells to approximately twice the level of unstimulated or propranolol-inhibited controls. The RIA appears to offer promising new approaches for studies on mucin metabolism and secretion in health and disease.