Matsumoto F S, Oliveira V C, Mançanares C A F, Ambrosio C E, Miglino M A
Laboratório de Anatomia de Animais Domésticos e Silvestres, Departamento de Cirurgia, Faculdade de Medicina Veterinária e Saúde Animal, Universidade de São Paulo, São Paulo, SP, Brasil.
Genet Mol Res. 2012 Nov 14;11(4):3942-54. doi: 10.4238/2012.November.14.1.
The yolk sac is an embryonic membrane that is essential for the embryo's initial survival in many mammals. It also plays an important role in the production of proteins necessary for development. We studied proteins of the yolk sac in bovine embryos at up to 40 days of gestation. We examined the yolk sac of 17 bovine embryos at different gestational periods, measuring α-fetoprotein, α-1-antitrypsin, and transferrin. This experiment was carried out by Western blot technique, associated with electrophoresis on a 6% sodium dodecyl sulfate polyacrylamide gel. Mouse monoclonal antibody anti-human-α-fetoprotein, mouse antibody anti-human-transferrin and rabbit polyclonal anti-human-α-1-antitrypsin were used as primary antibodies, and conjugated peroxidase as a secondary antibody. We detected the three proteins in some of the yolk sac samples; however, the bands in some specimens (samples) were weak, maybe a result of poor antigen-antibody reaction, since the antibodies used in this study were not specific to bovine proteins. The fact that weak bands appeared might be due to a weak cross-reaction.
卵黄囊是一种胚胎膜,对许多哺乳动物胚胎的早期存活至关重要。它在产生发育所需蛋白质方面也起着重要作用。我们研究了妊娠40天内牛胚胎的卵黄囊蛋白质。我们检查了17个处于不同妊娠期的牛胚胎的卵黄囊,检测甲胎蛋白、α-1抗胰蛋白酶和转铁蛋白。本实验采用蛋白质印迹技术,结合在6%十二烷基硫酸钠聚丙烯酰胺凝胶上进行电泳。小鼠抗人甲胎蛋白单克隆抗体、小鼠抗人转铁蛋白抗体和兔抗人α-1抗胰蛋白酶多克隆抗体用作一抗,共轭过氧化物酶用作二抗。我们在一些卵黄囊样本中检测到了这三种蛋白质;然而,一些标本(样本)中的条带较弱,这可能是抗原-抗体反应不佳的结果,因为本研究中使用的抗体并非牛蛋白质特异性抗体。出现弱条带这一事实可能是由于交叉反应较弱。