BMP-9 Antibody

R&D Systems | Catalog # AF3209

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Applications

Validated:

Neutralization

Cited:

Immunohistochemistry, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all BMP-9 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human BMP-9
Ser320-Arg429
Accession # Q9UK05

Specificity

Detects human BMP-9 in direct ELISAs. In direct ELISAs, 100% cross-reactivity with recombinant mouse BMP‑9 is observed. In direct ELISAs less than 2% cross-reactivity with recombinant human (rh) BMP-2, -3, -4, -5, -7, -8, andrhBMP‑10 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for BMP-9 Antibody

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

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

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.
Detection of BMP-9 by Western Blot

Detection of BMP-9 by Western Blot

HCMV US18 and US20 inhibit BMP9-mediated signaling during infection.(A) Schematic representation of the workflow. Recombinant HCMV US18stop, HCMV US20stop, and HCMV US18/20stop were constructed by introducing a 16 base pair (bp) stop cassette within the respective coding region. HFF-1 were infected by centrifugal enhancement with HCMV WT, HCMV US18stop, HCMV US20stop, or HCMV US18/20stop (MOI 4 for the 3 h time point, MOI 0.5 for the 48 h time point). Three or 48 h post infection, cells were stimulated with BMP9 (3 nM) for (1) 2 h, followed by cell lysis and immunoblot analysis, or (2) 6 h, followed by RNA extraction from cell lysates and RT-qPCR. (B, C) Cell lysates from (A) were subjected to immunoblot analysis with p-SMAD1/5/9, SMAD1, HCMV UL35, and GAPDH-specific antibodies (top panel), and transcript levels of Id3 and Stat2 are shown below. Data information: (B, 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 < 0.001, ****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.
Detection of BMP-9 by Western Blot

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.
Detection of BMP-9 by Western Blot

Detection of BMP-9 by Western Blot

HCMV US18 and US20 inhibit BMP9-mediated signaling during infection.(A) Schematic representation of the workflow. Recombinant HCMV US18stop, HCMV US20stop, and HCMV US18/20stop were constructed by introducing a 16 base pair (bp) stop cassette within the respective coding region. HFF-1 were infected by centrifugal enhancement with HCMV WT, HCMV US18stop, HCMV US20stop, or HCMV US18/20stop (MOI 4 for the 3 h time point, MOI 0.5 for the 48 h time point). Three or 48 h post infection, cells were stimulated with BMP9 (3 nM) for (1) 2 h, followed by cell lysis and immunoblot analysis, or (2) 6 h, followed by RNA extraction from cell lysates and RT-qPCR. (B, C) Cell lysates from (A) were subjected to immunoblot analysis with p-SMAD1/5/9, SMAD1, HCMV UL35, and GAPDH-specific antibodies (top panel), and transcript levels of Id3 and Stat2 are shown below. Data information: (B, 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 < 0.001, ****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.

Applications for BMP-9 Antibody

Application
Recommended Usage

Neutralization

Measured by its ability to neutralize BMP‑9-induced alkaline phosphatase production in the ATDC5 mouse chondrogenic cell line. The Neutralization Dose (ND50) is typically 0.005-0.1 µg/mL in the presence of 2 ng/mL Recombinant Human BMP‑9.

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: BMP-9

Human BMP-9, also known as growth and differentiation factor 2 (GDF-2), is a member of the BMP subgroup of the TGF-beta superfamily proteins that signal through heterodimeric complexes composed of type I and type II BMP receptors. BMP-9 regulates the development and function of a variety of embryonal and adult tissues (1, 2). The human BMP-9 cDNA encodes a 429 amino acid (aa) precursor that includes a 22 aa signal sequence, a 298 aa propeptide, and a 111 aa mature protein (3). Unlike with other BMP family proteins, the propeptide does not interfere with the biological activity of BMP-9 and remains associated with the mature peptide after proteolytic cleavage (4). Human and mouse BMP-9 share 96% aa sequence identity. Within the mature protein, human BMP-9 shares 64% aa sequence identity with human BMP-10 and less than 50% aa sequence identity with other BMPs. BMP-9 is expressed by non-parenchymal cells in the liver, (5, 6) where it promotes lipid metabolism and inhibits glucose production (7). BMP-9 exerts a prolonged hypoglycemic effect which may be due to an enhancement of insulin release (7). BMP-9 interacts with a high affinity specific heteromeric receptor expressed on liver endothelial cells that has been identified as ALK-1 (4-6). In the embryonal CNS, BMP-9 functions in the development and maintenance of the cholinergic neuronal phenotype (8-10). BMP-9 also induces the differentiation of mesenchymal stem cells into the chondrogenic lineage (11, 12). At low concentrations, BMP-9 is a proliferative factor for hematopoietic progenitor cells, but at higher concentrations, it enhances TGF-beta 1 production and inhibits hematopoietic progenitor colony formation (13).

References

  1. Chen, D. et al. (2004) Growth Factors 22:233.
  2. Miyazono, K. et al. (2005) Cytokine Growth Factor Rev. 16:251.
  3. Celeste, A.J. et al. (1994) J. Bone Miner. Res. 9:S136.
  4. Brown, M.A. et al. (2005) J. Biol. Chem. 280:25111.
  5. Song, J.J. et al. (1995) Endocrinology 136:4293.
  6. Miller, A.F. et al. (2000) J. Biol. Chem. 275:17937.
  7. Chen, C. et al. (2003) Nat. Biotechnol. 21:294.
  8. Lopez-Coviella, I. et al. (2000) Science 289:313.
  9. Lopez-Coviella, I. et al. (2005) Proc. Natl. Acad. Sci. USA 102:6984.
  10. Lopez-Coviella, I. et al. (2002) J. Physiol. Paris 96:53.
  11. Majumdar, M.K. et al. (2001) J. Cell. Physiol. 189:275.
  12. Hills, R.L. et al. (2005) J. Orthoped. Res. 23:611.
  13. Ploemacher, R.E. et al. (1999) Leukemia 13:428.

Long Name

Bone Morphogenetic Protein 9

Alternate Names

BMP9, GDF-2, GDF2

Entrez Gene IDs

2658 (Human); 12165 (Mouse)

Gene Symbol

GDF2

UniProt

Q9UK05

Additional BMP-9 Products

Product Documents for BMP-9 Antibody

Certificate of Analysis

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Product Specific Notices for BMP-9 Antibody

For research use only

Related Research Areas

BMP Family
Glucose Homeostasis

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