BMP-9 Antibody

Alkaline Phosphatase Production Induced by BMP‑9 and Neutralization by Human BMP‑9 Antibody.

Recombinant Human BMP-9 (Catalog # 3209-BP) induces alkaline phosphatase production in the ATDC5 mouse chondrogenic cell line in a dose-dependent manner (orange line). Alkaline phospha-tase production elicited by Recombinant Human BMP-9 (2 ng/mL) is neutralized (green line) by increasing concen-trations of Goat Anti-Human BMP-9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3209). The ND₅₀ is typically 0.005-0.1 µg/mL.

Human BMP-9 ELISA Standard Curve

Recombinant Human BMP‑9 (Catalog # 3209-BP) was serially diluted and captured by Mouse Anti-Human/Mouse/Primate BMP‑9 Monoclonal Antibody (Catalog # MAB3209) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human/Mouse BMP‑9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3209) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)

Detection of BMP-9 by Western Blot

BMP9 enhances STAT1 activation upon type I IFN, but not type II IFN stimulation.(A) After 2 h of serum starvation, HFF-1 were stimulated for either one or 8 h with IFN beta (5 ng/ml) or BMP9 (3 nM) alone, or were co-stimulated with IFN beta and BMP9, followed by cell lysis and immunoblot analysis with antibodies for phospho-STAT1, STAT1, phospho-SMAD1/5/9, and beta -Actin. Phospho-STAT1 band intensities were first normalized to corresponding total STAT1 levels, then to IFN beta stimulation only, and are shown below. (B) After 2 h of serum starvation, HFF-1 were stimulated for 1 h with either IFN alpha 2 (5 ng/ml), IFN gamma (5 ng/ml), or BMP9 (3 nM) alone, or co-stimulated with IFN alpha 2 or IFN gamma and BMP9, followed by cell lysis, immunoblot analysis and quantification as in (A). (C) After 2 h of serum starvation, HFF-1 were stimulated for 6 h with either IFN alpha 2 (5 ng/ml), IFN beta (5 ng/ml), IFN gamma (5 ng/ml), or BMP9 (3 nM) alone, or co-stimulated with IFN alpha 2, IFN beta, IFN gamma, and BMP9, followed by RNA extraction from cell lysates and RT-qPCR for Irf9, Stat2, Irf1, and Id3 transcripts. Data information: (A, B) Experiment was performed three independent times, one representative immunoblot is shown. Quantified data for the STAT1 phosphorylation levels are combined from three independent experiments. (C) Data are combined from three independent experiments. Student’s t test (unpaired, two-tailed), n.s. not significant, *P < 0.05, **P < 0.01, ****P < 0.0001. Data are shown as mean ± SD. Source data are available online for this figure. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/38308064), licensed under a CC-BY license. Not internally tested by R&D Systems.