A method has been described for the purification of ovine placental lactogen (oPL) involving the use of freshly obtained sheep foetal cotyledons. Tissue was extracted with 0·1 m-ammonium bicarbonate and the supernatant fraction, adjusted to pH 7, was brought to 60% saturation with ammonium sulphate. The resulting precipitate was then subjected to a sequence of chromatographic steps using columns of Sephadex G-100 and carboxymethylcellulose. During each stage of the purification, the lactogenic activity was monitored with a pregnant rabbit mammary gland radioreceptor assay. The yield of oPL corresponded to 8 mg/kg wet foetal tissue and the oPL possessed lactogenic activity equivalent to 1 mg ovine prolactin/mg protein and GH-like activity equivalent to 0·8 mg human GH/mg protein. The biological activity of oPL was confirmed using a rabbit intraductal mammary gland assay in vivo.
After polyacrylamide gel electrophoresis at pH 8·9, oPL was resolved into one major band (isoelectric point 8·2–8·4) and four minor components, which were thought to be deamidation products of oPL. Microimmunoelectrophoresis and immunodiffusion studies confirmed that the preparation of oPL was free from serum protein contaminants.
Journal of Endocrinology is committed to supporting researchers in demonstrating the impact of their articles published in the journal.
The two types of article metrics we measure are (i) more traditional full-text views and pdf downloads, and (ii) Altmetric data, which shows the wider impact of articles in a range of non-traditional sources, such as social media.
More information is on the Reasons to publish page.
Sept 2018 onwards | Past Year | Past 30 Days | |
---|---|---|---|
Full Text Views | 1 | 0 | 0 |
PDF Downloads | 2 | 1 | 0 |