Isolation, characterization and immunocytochemical localization of bovine trophoblast protein-1.
Biol Reprod
; 40(2): 343-52, 1989 Feb.
Article
em En
| MEDLINE
| ID: mdl-2720031
ABSTRACT
Bovine trophoblast protein-1 (bTP-1) was isolated to 90% purity from culture medium of Day 18-20 conceptuses incubated in vitro (in the presence of L-[3H]leucine) by a combination of Sephacryl S-200 gel filtration chromatography and O-(diethylaminoethyl) (DEAE) anion-exchange high-performance liquid chromatography (DEAE-HPLC). The radiolabeled protein had an Mr of 21,200 +/- 800 by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and had three isoelectric variants (pI 5.7-6.5) by two-dimensional SDS-PAGE. DEAE-HPLC-enriched bTP-1 cross-reacted with anti-o TP-1 serum on Western blots. A monospecific antiserum against bTP-1 was produced by excising the bTP-1 polypeptide band from preparative SDS-PAGE gels. Antiserum reacted with a single polypeptide with an Mr of 21,200 as determined by Western blotting of SDS-PAGE-separated conceptus medium proteins and by immunoprecipitation from L-[35S]methionine-labeled medium proteins followed by SDS-PAGE and autoradiography. Bovine trophoblast protein-1 was localized by immunocytochemistry in the cytoplasm of both mono- and binuclear trophectoderm cells of Day 20 bovine conceptuses, indicating that it is a product of the trophoblast.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proteínas da Gravidez
/
Interferon Tipo I
/
Embrião de Mamíferos
Limite:
Animals
/
Pregnancy
Idioma:
En
Revista:
Biol Reprod
Ano de publicação:
1989
Tipo de documento:
Article