The preparation of a highly purified antibody and a solid-state immunoassay for porcine pancreatic α-amylase

H. James Wedner, Lawrence N. Parker, Michael G. Rosenfeld

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Porcine pancreas and parotid cell suspensions provide model systems in which to study the mechanism of induction of a specific protein, α-amylase, by hormones acting via cAMP. A highly purified antibody against porcine pancreatic α-amylase was prepared using affinity chromatography of the total IgG fraction derived from rabbit anti-α-amylase antiserum and was used to develop a radioimmunoassay for α-amylase. The antigen-antibody complex was separated from unbound α-amylase using either glutaraldehyde gelled α-amylase or a second antibody technique; a linear standard curve was achieved over a 100- to 1000-fold range of α-amylase concentration. Tissue homogenates did not interfere with this assay, and assayed levels of α-amylase in porcine pancreas were linear using 10-200 μg of homogenate. No levels or very low levels of α-amylase were detected in control tissues.

Original languageEnglish
Pages (from-to)175-186
Number of pages12
JournalAnalytical Biochemistry
Volume65
Issue number1-2
DOIs
StatePublished - May 12 1975

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