bmpr2 fc  (R&D Systems)

Journal: BMC Research Notes

Article Title: Identification of a bone morphogenetic protein type 2 receptor neutralizing antibody

doi: 10.1186/s13104-019-4367-0

Figure Lengend Snippet: Antibody 3F6 reduces BMPR2-ECD ligand-binding activity in a modified immunoprecipitation assay. Neutralizing the ligand-binding activity of BMPR2-ECD using various amounts of 3F6. Results are quantified by ELISA and expressed as mean ± SEM relative to the ligand-binding activity of BMPR2-ECD in the absence of 3F6. n ≥ 3 per condition. Asterisk indicates p < 0.05 by paired t test

Article Snippet: BMPR2-ECD/Fc fusion (Sino Biologicals 10551-H03H) was mixed with 5 µL Protein G-coupled Dynabeads (Invitrogen 1003D) at room temperature for 30 min in 200 µL total volume with gentle rocking.

Techniques: Ligand Binding Assay, Activity Assay, Modification, Immunoprecipitation, Enzyme-linked Immunosorbent Assay

Journal: BMC Research Notes

Article Title: Identification of a bone morphogenetic protein type 2 receptor neutralizing antibody

doi: 10.1186/s13104-019-4367-0

Figure Lengend Snippet: Antibody 3F6 reduces activation of BMP pathway in HEK293T cells. a Expression of endogenous BMPR2 by HEK293T cells compared to β-actin loading control. Approximate molecular weights are indicated. b , c BMP2 induces phosphorylation of SMAD1, 5, and 8 (pSMAD1,5,8) in HEK293T cells and this response is blunted by pre-treatment with 3F6. Approximate molecular weights are indicated in b . Results are quantified in c and expressed as mean ± SEM ratio of phosphorylated SMAD1,5,8: total SMAD1 relative to BMP2 treatment alone. No effect on BMP2-responsiveness was observed with pre-treatment using control ascites (Ctrl Asc.). n = 3 per condition. Asterisk indicates p < 0.05 by paired t test

Article Snippet: BMPR2-ECD/Fc fusion (Sino Biologicals 10551-H03H) was mixed with 5 µL Protein G-coupled Dynabeads (Invitrogen 1003D) at room temperature for 30 min in 200 µL total volume with gentle rocking.

Techniques: Activation Assay, Expressing

Journal: BMC Research Notes

Article Title: Identification of a bone morphogenetic protein type 2 receptor neutralizing antibody

doi: 10.1186/s13104-019-4367-0

Figure Lengend Snippet: Antibody 3F6 has no effect on BMP-responsiveness in BMPR2 knock-down HEK293T cells. a Expression levels of endogenous BMPR2 in scramble control HEK293T cells (Control) and HEK293T cells carrying anti- BMPR2 shRNA ( BMPR2 -KD) compared to HPRT1 loading control. Data are expressed as normalized to scramble control (Control) using the 2 −∆∆Ct method. n = 6 per condition. Asterisk indicates p < 0.05 by unpaired t test. b , c Expression of endogenous BMPR2 in scramble control HEK293T cells (Control) and BMPR2 -KD HEK293T cells compared to β-actin loading control. Approximate molecular weights are indicated in b . Representative immunoblot is shown in b and results from three independent runs are quantified in c (data are expressed as mean ± SEM ratio of BMPR2: β-actin normalized to scramble control (Relative Expression)). Asterisk indicates p < 0.05 by paired t test. d , e BMP2 induces phosphorylation of SMAD1, 5, and 8 (pSMAD1,5,8) in BMPR2 - KD HEK293T cells. No effect on BMP2-responsiveness of BMPR2 - KD HEK293T cells was observed with pre-treatment using control ascites (Ctrl Asc.) or 3F6. Approximate molecular weights are indicated in d . Results are quantified in E and expressed as mean ± SEM ratio of phosphorylated SMAD1,5,8: total SMAD1 relative to BMP2 treatment alone (Relative pS1:S1). n = 4 per condition

Article Snippet: BMPR2-ECD/Fc fusion (Sino Biologicals 10551-H03H) was mixed with 5 µL Protein G-coupled Dynabeads (Invitrogen 1003D) at room temperature for 30 min in 200 µL total volume with gentle rocking.

Techniques: Expressing, shRNA, Western Blot