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Generation and characterization of a polyclonal antipeptide antibody to human glycodelin
A S Poddar1, J G Kim, K P Gill
1Department of Gynecology and Obstetrics, Emory University, Atlanta, Georgia 30322, USA.
Objective:
To develop and characterize an antiglycodelin antibody using a 15-amino acid synthetic peptide as antigen, derived from the sequence of human glycodelin.
Design:
We have developed a chicken antiglycodelin-derived peptide antibody and have characterized the antibody with the use of endometrial and ovarian cell lines. The antibody was also tested for its ability to detect glycodelin by ELISA assay, immunocytochemistry, and by Western blot.
Setting:
Various cell lines, cell culture medium, and amniotic fluid were used in the experiments.
Patient(S):
Amniotic fluid was collected from pregnant patients in their first trimester of pregnancy.
Intervention(S):
No intervention.
Main Outcome Measure(S):
Detection of glycodelin.
Result(S):
The cell lines RL95-2 (human endometrial carcinoma cells), OVCAR-3 (human ovarian adenocarcinoma cells), HeLa (human cervical epitheloid carcinoma cells), and EM42-D (human endometrial epithelial cells) reacted with the antibody, indicating the presence of glycodelin. A specific 45-kd protein representing glycodelin was detected by Western blot in the amniotic fluid.
Conclusion(S):
Antipeptide antibodies can be successfully used to detect and quantify the presence of glycodelin in cells and fluids.