lipolysis bmprii fc human mouse 811 br 100 r d systems spr  (R&D Systems)

Journal: The FASEB Journal

Article Title: Prodomain processing controls BMP‐10 bioactivity and targeting to fibrillin‐1 in latent conformation

doi: 10.1096/fj.202401694r

Figure Lengend Snippet: FIGURE 4 The processed BMP-10 CPLX binds robustly to BMP receptors whereas the unprocessed dimer shows no receptor interaction. Sensorgrams of SPR interaction studies of soluble BMP-10 GF, processed BMP-10 CPLX, and unprocessed BMP-10 dimer flowed over immobilized BMP receptors. Soluble analytes were injected onto immobilized BMPRII, ALK-1, and ENG at concentrations ranging from 0 to 80 nM. KDs were calculated from three independent experiments (N = 3).

Article Snippet: To assess the binding affinity of BMP- 10 processing variants to BMP receptors, the human IgG1- Fc- fusion ectodomains of BMPRII (#811- BR- 100/CF, R&D Systems), ALK- 1 (#370- AL- 100/CF, R&D Systems) as well as ENG (#1097- EN- 025/CF, R&D Systems) were immobilized at 500 or 800 RUs on a CM5 chip via amine coupling.

Techniques: Injection

Journal: Biomaterials advances

Article Title: Enhancing BMP-2-mediated osteogenesis with a synthetic heparan sulfate mimetic.

doi: 10.1016/j.bioadv.2023.213671

Figure Lengend Snippet: Fig. 2. Glycomimetic 15 binds BMP-2 and several other pro-regenerative factors with high affinity. A – SPR solution competition dosing experiments were performed with Ai - BMP-2, Aii - BMP-4, Aiii - BMP-7, Aiv BMPRII Fc chimera and Av - VEGF165 using a dose range of glycomimetic 15 (3.91 to 8000 nM). Avi – normalized response data extrapolated from sensorgrams Ai-Av. B – fluorescence polarization solution competition with BMP-2 (50 nM), 488-labelled heparin dp24 (0.5 nM) and increasing concentrations of glycomimetic 15 (red) or glycomimetic 18 (blue). Ci – DSF melt curves. Cii – first derivative of melt curve data. Ciii – BMP-2 melt temperature when incubated with multiple concentrations of glycomimetic 15, derived from the first derivative of melt curve data. Di – Normalized anti-factor Xa activity for glycomimetic 15 (black), heparin dp12 (red) and heparin dp24 (blue). Dii – anti-factor Xa activity in IU/mg, calculated using USP heparin standard curve. Bars represent mean, and error bars represent S.D. All data is representative of three independent experiments.

Article Snippet: BMP-2 (cat. # 355-BM/ CF), BMP-4 (cat. # 314-BP/CF), BMP-7 (cat. # 354-BP/CF), BMPRII Fc chimera (cat. # 811-BR), Noggin (cat. # 6057-NG/CF), VEGF165 (cat. # 293-VE/CF) polyclonal Goat anti-Human BMP-2/BMP-4 IgG (cat. # AF355), biotinylated monoclonal Mouse anti-Human BMP-2 IgG (cat. #BAM3552) were purchased from R&D systems.

Techniques: Fluorescence, Incubation, Derivative Assay, Activity Assay

Journal: Biomaterials advances

Article Title: Enhancing BMP-2-mediated osteogenesis with a synthetic heparan sulfate mimetic.

doi: 10.1016/j.bioadv.2023.213671

Figure Lengend Snippet: Fig. 5. Glycomimetic 15 influences BMPRII: BMP-2 and BMP-2: cell surface interactions. Ai – Co-immunoprecipitation and Western blot of BMP-2 (125 nM) with BMPRII Fc chimera (250 nM) ± increasing concentrations of either glycomimetic 15 or 18. Aii – Log2 fold change of densitometry values obtained from three in dependent co-immunoprecipitation experiments – bars represent mean log2 fold change ±95 % CI, normalized to BMPRII + BMP-2 alone. Bi – Flow cytometry histograms of C2C12 cells stained with BMP-2 (black histogram) ± increasing concentrations of glycomimetic 15 (red histogram) or glycomimetic 18 (blue histo gram). Bii – Cell surface binding of BMP-2 to C2C12 cells incubated with increasing concentrations of glycomimetic 15 (red) or glycomimetic 18 (blue) – data is mean geometric mean fluorescence ± S.D. normalized to BMP-2 alone. All data either constitutes (Aii, Bii) or is representative of (Ai, Bi) three independent experiments. Statistical analysis was performed via one-way ANOVA with Tukey’s post hoc testing (Aii) or two-way ANOVA with ˇSíd´ak or Bonferroni post hoc testing (Bii). Sig nificance is denoted by *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001.

Article Snippet: BMP-2 (cat. # 355-BM/ CF), BMP-4 (cat. # 314-BP/CF), BMP-7 (cat. # 354-BP/CF), BMPRII Fc chimera (cat. # 811-BR), Noggin (cat. # 6057-NG/CF), VEGF165 (cat. # 293-VE/CF) polyclonal Goat anti-Human BMP-2/BMP-4 IgG (cat. # AF355), biotinylated monoclonal Mouse anti-Human BMP-2 IgG (cat. #BAM3552) were purchased from R&D systems.

Techniques: Immunoprecipitation, Western Blot, Flow Cytometry, Staining, Binding Assay, Incubation, Fluorescence