276. The technical and biological validation of an LH assay for use with the Western Grey Kangaroo (Macropus fuliginosis) and Black-flanked Rock Wallaby (Petrogale lateralis lateralis)
Reproduction, Fertility and Development(2008)
摘要
Methods for the measurement of marsupial LH invariably rely upon the similarity of the LH molecule between different species and usually use anti-ovine or anti-bovine LH antibody and an ovine or bovine labelled LH preparation. Initial attempts to measure plasma LH in the Western Grey Kangaroo with assays using antibodies to 4 different isoforms of ovine LH raised in 7 different rabbits were unsuccessful. An enzymeimmunoassay (EIA) developed for the Asian elephant (Zoo Biology 23:45–63) was then applied to the Western Grey Kangaroo and the Black-flanked Rock Wallaby. This EIA has an anti-bovine-LH monoclonal antibody (518B7 provided by Dr Jan Roser, University of California, Davis, USA), biotinylated ovine LH label and bovine LH standard (NIADDK-oLH-26 and NIH-bLH-B10, both provided by Dr Janine Brown and Nicole Abbondanza, Smithsonian Institute, Front Royal, Virginia USA). Technical validation showed that serial dilution down to 1:8 of plasma from 7 individuals of each species showed parallelism to the assay standard curve, and control samples (1.24–5.30 ng/mL) had between-assay coefficients of variation 0.0005) were seen after GnRH for both the Western Grey Kangaroo (from 5.0 ± 0.8 ng/mL to 9.4 ± 1.2 ng/mL; n = 19) and the Black-flanked Rock Wallaby (from 6.0 ± 0.7 ng/mL to 10.6 ± 0.6 ng/mL; n = 28). In conclusion, this assay can be successfully used to measure LH in these two species.
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transgenesis,andrology,embryonic development,fertility,embryogenesis,embryo development,theriogenology,gametogenesis,art,cryopreservation,folliculogenesis,ivf,reproductive biology,embryo,oogenesis,genetics,cell biology,educational,endocrinology,fertilization,fertilisation,cloning,spermatogenesis
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