Anti-20-alpha-Hydroxyprogesterone polyclonal antibody

In the present work we optimised the separation of battery of key UV non-transparent low-molecular-mass compounds having possible endocrine disrupting compounds (EDCs) activity or which may be used as the endocrine effect biomarkers. Simple Optimization strategy was based oil strong temperature effect that is driven by electrostatic interactions between macrocyclic mobile phase additives like cyclodextrins and eluted components of interest under C18 stationary phase and acetonitrile/water mobile phase conditions. Particularly. the effect of temperature involving native beta-cyclodextrin and its hydroxypropyl derivative to improve separation of number of natural (d-equilenin, equilin, estetrol, estriol, estrone, 17 beta-estradiol, 17 alpha-hydroxyprogesterone, 20 alpha-hydroxyprogesterone, cortisol. cortisone, progesterone, testosterone, tetrahydrocortisol and tetra hydrocortisone) and artificial steroids (ethynylestracliol, norgestrel isomers, medroxyprogesterone, mestranol. methyltestosterone, norethindrone, 17 alpha-estradiol) as well as non-steroidal compounds (diethylstilbesterol, bisphenol A. 4-tert-butylphenol, dimethyl phthalate, dibutyl phthalate and dioctyl phthalate) was investigated. It has been found that successful isocratic separation of 27 chemicals call be achieved using acetonitrile/water eluents modified with beta-cyclodextrin or hydroxypropyl-beta-cyclodextrin at concentration of 10 mM and temperature e of 47 C Separation protocol is simple, reliable, direct and non-radioactive and may be easily adapted for rapid separation and quantification of wide range of given steroids and related EDCs in environmental samples, particularly those that are characterised by unstable biological matrix and components of interest load (C) 2009 Elsevier B.V. All rights reserved.

The enzyme 20 alpha-hydroxysteroid dehydrogenase (20 alpha-HSD) catalyzes the conversion of progesterone to its inactive form, 20 alpha-hydroxyprogesterone. This enzyme plays a critical role in the regulation of luteal function in female mammals. In this study, we conducted the characterization and functional analyses of bovine 20 alpha-HSD from placental and ovarian tissues. The nucleotide sequence of bovine 20 alpha-HSD showed significant homology to that of goats (96%), humans (84%), rabbits (83%), and mice (81%). The mRNA levels increased gradually throughout the estrous cycle, the highest being in the corpus luteum (CL) 1 stage. Northern blot analysis revealed a 1.2 kb mRNA in the bovine placental and ovarian tissues. An antibody specific to bovine 20 alpha-HSD was generated in a rabbit immunized with the purified, recombinant protein. Recombinant 20 alpha-HSD protein produced in mammalian cells had a molecular weight of similar to 37 kDa. Bacterially expressed bovine 20 alpha-HSD protein showed enzymatic activity. The expression pattern of the 20 alpha-HSD protein in the pre-parturition placenta and the CL1 stage of the estrous cycle was similar to the level of 20 alpha-HSD mRNA expression. Immunohistochemical analysis also revealed that bovine 20 alpha-HSD protein was intensively localized in the large luteal cells during the late estrous cycle. Reproduction (2011) 142 723-731