caa a22 4500 465804543 CHVBK 20180323112100.0 cr unu---uuuuu 170327e19900301xx s 000 0 eng 10.1007/BF03349541 doi (NATIONALLICENCE)springer-10.1007/BF03349541 Extraction of rat pituitary prolactin for radioimmunoassay [Elektronische Daten] [A. Torres, M. Prada, A. Aoki] The bulk of Prolactin (PRL) in rat pituitary gland stored as big molecular forms in secretory granules is mostly excluded from radioimmunoassay (RIA) determinations because secretory granules remain intact after tissue homogenization. Also big PRL is little immunoreactive and must be deaggregated to monomers to allow a complete detection by RIA. Different dissociating agents, used to render PRL monomers soluble, were tested at various temperatures and pH conditions. Incubations of pituitary homogenates in buffers at neutral pH yield consistent levels of PRL, but the sole alkalinization of the media increases significantly the radioimmunoassayable PRL content. No significant increase was detected with EDTA and thiols. The 2.5 M urea was the most effective extraction agent increasing about 7-fold the quantification of PRL by RIA. Extraction of PRL with urea was enhanced at pH 9.0 and at 4 C and this combination constitutes the method of choice for a complete extraction of pituitary PRL. Italian Society of Endocrinology (SIE), 1990 Prolactin nationallicence extraction nationallicence RIA nationallicence pituitary gland nationallicence PRL polymers nationallicence Torres A. Centro de Microscopia Electronica, Facultad de Ciencias Medicas, Universidad Nacional de Cordoba, Casilla Postal 362, 5000, Cordoba, Argentina aut Prada M. Centro de Microscopia Electronica, Facultad de Ciencias Medicas, Universidad Nacional de Cordoba, Casilla Postal 362, 5000, Cordoba, Argentina aut Aoki A. Centro de Microscopia Electronica, Facultad de Ciencias Medicas, Universidad Nacional de Cordoba, Casilla Postal 362, 5000, Cordoba, Argentina aut Journal of Endocrinological Investigation Springer International Publishing 13/3(1990-03-01), 205-208 0391-4097 13:3<205 1990 13 40618 https://doi.org/10.1007/BF03349541 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF03349541 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Torres A. Centro de Microscopia Electronica, Facultad de Ciencias Medicas, Universidad Nacional de Cordoba, Casilla Postal 362, 5000, Cordoba, Argentina aut NATIONALLICENCE 700 1- Prada M. Centro de Microscopia Electronica, Facultad de Ciencias Medicas, Universidad Nacional de Cordoba, Casilla Postal 362, 5000, Cordoba, Argentina aut NATIONALLICENCE 700 1- Aoki A. Centro de Microscopia Electronica, Facultad de Ciencias Medicas, Universidad Nacional de Cordoba, Casilla Postal 362, 5000, Cordoba, Argentina aut NATIONALLICENCE 773 0- Journal of Endocrinological Investigation Springer International Publishing 13/3(1990-03-01), 205-208 0391-4097 13:3<205 1990 13 40618 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer