TY - JOUR
T1 - The preparation of a highly purified antibody and a solid-state immunoassay for porcine pancreatic α-amylase
AU - Wedner, H. James
AU - Parker, Lawrence N.
AU - Rosenfeld, Michael G.
N1 - Funding Information:
The authors gratefully acknowledge the aid of Dr. Gary David for iodination techniques and Dr. Charles Parker for advice and review of the manuscript. This investigation was supported by PHS Research Grant No. AM13 149 from the National Institute of Arthritis, Metabolism & Digestive Diseases and by Grant No. BC169 from the American Cancer Society.
PY - 1975/5/12
Y1 - 1975/5/12
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0016850033&partnerID=8YFLogxK
U2 - 10.1016/0003-2697(75)90503-5
DO - 10.1016/0003-2697(75)90503-5
M3 - Article
C2 - 1130675
AN - SCOPUS:0016850033
SN - 0003-2697
VL - 65
SP - 175
EP - 186
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 1-2
ER -