Evaluación de la especificidad de anticuerpos policlonales aviarios y mamíferos anti-Factor Precoz de Preñez porcino

Grosso, M.; Cuello, M.F.; Martínez, Rosario; Greco, Claudio; Henklein, Peter; Schade, Rüdiger; Vivas, Adriana

doi:10.1016/s0213-9626(08)70044-1

Cited by 2 publications

(5 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We produced polyclonal antibodies to synthetic peptides that corresponded to different parts of the amino acid sequence of EPF to produce a more robust array of reagents (Grosso et al 2008).…”

Section: Tissue Samplingmentioning

confidence: 99%

“…In a study of rat tissues, using electron microscopy and polyclonal antibodies, Sadacharan et al (2001) found EPF in extramitochondrial compartments. EPF localization studies of human colon carcinoma tissue showed diffuse punctate cytoplasmic EPF in reproductive tissues of pregnant gilts 3 standard (Grosso et al 2008). The proteins were separated electrophoretically according to Laemmli (1970) and analyzed by western immunoblot by probing with rabbit polyclonal anti-porcine EPF (1:100 in 10% skim milk in TBS-T) overnight at 4 ° C as the primary antibody, and peroxidase-conjugated anti-rabbit immunoglobulin as the secondary antibody (A6154; Sigma) 1:1000 in 10% skim milk in TBS-T for 90 min at room temperature.…”

Section: Tissue Samplingmentioning

confidence: 99%

“…Rabbit polyclonal anti-porcine EPF antibodies (IgG) developed in our laboratory (Grosso et al 2008) were used to determine the presence of EPF in serum and PPCM by western blot and in reproductive tissues by immunohistochemistry.…”

Section: Epf Determinationmentioning

confidence: 99%

“…All reproductive tissues of pregnant animals at all gestational stages analyzed showed specifi c EPF labeling, but no labeling was observed in tissues from nonpregnant gilts. (Grosso et al 2008) or pre-immune serum to controls. In the fi nal step, DAB (Sigma) was added as chromogen and all sections were counterstained with Mayer ' s hematoxylin.…”

Section: Epf In Reproductive Tissuesmentioning

confidence: 99%

“…The anti-EPF antibodies used for immunohistochemistry are specifi c against EPF obtained from swine serum at 7 days of gestation, so the antibodies would be directed against EPF produced by maternal tissues (ovary and oviduct) prior to implantation (Grosso et al 2008). These antibodies recognize and bind specifi cally to the EPF produced by the ovary and oviduct (pre-implantation) and to that produced by the embryo (post-implantation).…”

Section: Sixty Days Of Pregnancymentioning

confidence: 99%

See 4 more Smart Citations

Immunohistochemical distribution of early pregnancy factor in ovary, oviduct and placenta of pregnant gilts

Grosso¹,

Bellingeri²,

Motta³

et al. 2014

Biotechnic & Histochemistry

View full text Add to dashboard Cite

Early pregnancy factor (EPF) is an immunosuppressant that promotes maternal immune system tolerance of the allogenic fetus. Little is known about localization of this factor in different tissues and nothing has been reported about localization in swine reproductive and placental tissues. We determined the concentration of EPF in serum of gilts and porcine placenta conditioned medium (PPCM). We also analyzed the expression of EPF in different reproductive tissues of pregnant gilts at 10, 30, 60 and 90 days of pregnancy. EPF concentration in serum and PPCM was determined by western blot and densitometry. EPF expression in reproductive tissue was assessed by immunohistochemistry. The highest concentration of EPF was observed at 30 days in serum and PPCM; the concentration was higher in PPCM than in serum at the stages we evaluated. All reproductive tissues from the gestational stages analyzed showed specific labeling of EPF, but this labeling did not appear in non-pregnant gilts. At 30 days pregnancy, the EPF expression in the ovary was predominantly in follicular lutein cells, probably owing to its function as a luteotrophic factor. In the oviduct, EPF was expressed in unciliated secretory epithelial cells and in the cilia of ciliated cells. In the placenta, EPF was expressed in the fetal portion (mesoderm chorioallantois and epithelium of endoderm). EPF acts as an autocrine and paracrine growth factor for the trophoblast during the peri-implantation period.

show abstract

Section: Tissue Samplingmentioning

confidence: 99%