igg tgfb1 neutralizing antibody  (R&D Systems)

Journal: Biology of Reproduction

Article Title: Human Pregnancy Specific Beta-1-Glycoprotein 1 (PSG1) Has a Potential Role in Placental Vascular Morphogenesis ¹

doi: 10.1095/biolreprod.109.082412

Figure Lengend Snippet: PSG1 induces TGFB1 and VEGFA in human monocytes. Human monocytes were treated with increasing concentrations of PSG1-Fc or FLAG-Fc for 24 h. TGFB1 ( A ) and VEGFA ( B ) in the supernatants were analyzed by ELISA. All cells were treated in triplicate, and data are representative of at least three independent experiments. Statistical significance was determined by one-way ANOVA (* P < 0.01). Error bars the SEM.

Article Snippet: Neutralization of TGFB1 was performed by adding 10 μg/ml of the anti-human TGFB1 neutralizing antibody (R&D Systems) to the cells 10 min before PSG1 addition.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Biology of Reproduction

Article Title: Human Pregnancy Specific Beta-1-Glycoprotein 1 (PSG1) Has a Potential Role in Placental Vascular Morphogenesis ¹

doi: 10.1095/biolreprod.109.082412

Figure Lengend Snippet: PSG1 induces TGFB1 and VEGFA in human extravillous trophoblast cell lines. HTR-8/SVneo and SGHPL-4 cells were treated with different concentrations of PSG1-Fc ( A and C – E ) or PSG1-FLAG ( B ) for 24 h. TGFB1 and VEGFA in the supernatants were analyzed by ELISA. In C , HTR-8/SVneo cells were pre-treated with 10 μg/ml of anti-TGFB1 antibody or isotype-matched control antibody (IM) 1 h before addition of 50 μg/ml of PSG1-Fc or FLAG-Fc. After 24 h, VEGFA in the supernatants was analyzed by ELISA. All cells were treated in triplicate, and data are representative of at least three independent experiments. For A , B , D , and E , statistical significance was determined by one-way ANOVA, and the P -value between doses was less than 0.01. For C , statistical significance between the PSG1-Fc-treated and FLAG-Fc-treated samples was determined by two-tailed Student t -test (* P < 0.05). Error bars represent the SEM.

Article Snippet: Neutralization of TGFB1 was performed by adding 10 μg/ml of the anti-human TGFB1 neutralizing antibody (R&D Systems) to the cells 10 min before PSG1 addition.

Techniques: Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Biology of Reproduction

Article Title: Human Pregnancy Specific Beta-1-Glycoprotein 1 (PSG1) Has a Potential Role in Placental Vascular Morphogenesis ¹

doi: 10.1095/biolreprod.109.082412

Figure Lengend Snippet: PSG1 induces TGFB1 in human endothelial cells. UtMVECs ( A ) and HEEC ( B ) were treated with different concentrations of PSG1-Fc or FLAG-Fc. TGFB1 in the supernatants was measured by ELISA. All cells were treated in triplicate, and data are representative of at least three independent experiments. Statistical significance was determined by one-way ANOVA (* P < 0.01). Error bars represent the SEM.

Article Snippet: Neutralization of TGFB1 was performed by adding 10 μg/ml of the anti-human TGFB1 neutralizing antibody (R&D Systems) to the cells 10 min before PSG1 addition.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Biology of Reproduction

Article Title: Human Pregnancy Specific Beta-1-Glycoprotein 1 (PSG1) Has a Potential Role in Placental Vascular Morphogenesis ¹

doi: 10.1095/biolreprod.109.082412

Figure Lengend Snippet: The GDD residues of PSG1 are not required for activity while N -linked glycosylation of the N-domain is essential. A ) PSG1-Fc (lane 1), PSG1 gdd→sdl Fc (lane 2), and PSG1 rnn→aaa (lane 3) secreted by CHO cells were purified on a protein A column, separated on SDS-PAGE, and stained with GelCode Blue. B ) HTR-8/SVneo cells were treated with 30 μg/ml of PSG1-Fc, PSG1 gdd→sdl Fc, PSG1 rnn→aaa , and FLAG-Fc for 24 h. TGFB1 in the supernatants was analyzed by ELISA. All cells were treated in triplicate, and data are representative of at least three independent experiments. Statistical significance between the protein-treated and control-treated samples was determined by two-tailed Student t -test (* P < 0.05). Error bars represent the SEM.

Article Snippet: Neutralization of TGFB1 was performed by adding 10 μg/ml of the anti-human TGFB1 neutralizing antibody (R&D Systems) to the cells 10 min before PSG1 addition.

Techniques: Activity Assay, Purification, SDS Page, Staining, Enzyme-linked Immunosorbent Assay, Two Tailed Test