Rhbmp 7

Manufactured by R&D Systems

Sourced in United States

RhBMP-7 is a recombinant human bone morphogenetic protein-7 (BMP-7). BMP-7 is a member of the transforming growth factor-beta (TGF-beta) superfamily of proteins and plays a role in bone and cartilage development.

Automatically generated - may contain errors

Lab products found in correlation

Rhfgf9, by R&D Systems (3 mentions) Rhbmp 2, by R&D Systems (2 mentions) Rhbmp 4, by R&D Systems (2 mentions) Slide a lyzer dialysis cassette, by Thermo Fisher Scientific (1 mentions) Rhbmp 7, by ProSpec (1 mentions) Anti β tubulin, by Santa Cruz Biotechnology (1 mentions) Anti p jnk, by Cell Signaling Technology (1 mentions) Click it edu alexa fluor 488 imaging kit, by Thermo Fisher Scientific (1 mentions) Anti phh3, by Cell Signaling Technology (1 mentions) Hoechst 33342, by Thermo Fisher Scientific (1 mentions) Hydrochloric acid (hcl), by Kanto Chemical (1 mentions) Bovine serum albumin (bsa), by Thermo Fisher Scientific (1 mentions) Rhgrem1, by R&D Systems (1 mentions)

6 protocols using rhbmp 7

Optimization of rhBMP-7 Formulation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Prior to the in vivo studies, the biological activity of rhBMP-7 from different manufacturers was determined in an alkaline phosphatase (ALP) activity assay in ATDC5 cells, amongst them rhBMP-7 from R&D and ProSpec-Tany. rhBMP-7 from ProSpec-Tany showed the highest biological activity, as shown in Additional file 2, and was further chosen to be employed in the in vivo studies.
rhBMP-7 (CYT-276, ProSpec-Tany TechnoGene Ltd) was reconstituted in 60 μl sterile water. This solution was dialyzed against a buffer solution containing 17% sucrose, 20% mannitol, 332 mM glycine, and 0.8% Tween 20, using a slide-A-Lyzer dialysis cassette (66454, Thermo Fisher Scientific Inc., Rockford, IL, USA) with a molecular weight cutoff of 10 kDa overnight with 4 buffer changes. The final solution containing 300 μg of rhBMP-7 (calculated amount) was freeze-dried overnight and reconstituted in sterile water, to achieve a final concentration of 250 μg rhBMP-7 in 40 μl buffer solution containing 1% sucrose, 1.2% mannitol, 20 mM glycine, and 0.05% Tween 20. Activity of the dialysate was shown to be retained in vitro through its capacity to induce ALP production in mice ATDC5 cells. Additional file 2 shows the biologic activity of the dialyzed rhBMP-7 compared with the pre-dialyzed rhBMP-7.

Willems N., Bach F.C., Plomp S.G., van Rijen M.H., Wolfswinkel J., Grinwis G.C., Bos C., Strijkers G.J., Dhert W.J., Meij B.P., Creemers L.B, & Tryfonidou M.A. (2015). Intradiscal application of rhBMP-7 does not induce regeneration in a canine model of spontaneous intervertebral disc degeneration. Arthritis Research & Therapy, 17(1), 137.

+ Open protocol

+ Expand

BMP-mediated Smad1/5/8 Phosphorylation

Check if the same lab product or an alternative is used in the 5 most similar protocols

For BMP stimulation of Smad1/5/8 phosphorylation, HK-2 and HEK293 cells were plated on 60 mm plates. At 70 % confluence, cells were washed with 1 x PBS and treated with HK-2 complete medium supplemented with vehicle (4 mM HCl) or increasing concentrations of rhBMP-2, rhBMP-4 (0.5-10 ng/ml) or rhBMP-7 (5-50 ng/ml) (R&D Systems, Minneapolis, USA) for 60 min. For rhGrem1 inhibition experiments, HK-2 cells were treated with complete medium supplemented with vehicle (4 mM HCl), 5 ng/ml rhBMP-2, 5 ng/ml rhBMP-4 or 20 ng/ml rhBMP-7 in the absence or presence of increasing concentrations of rhGrem1 (5-400 ng/ml) (R&D Systems, Minneapolis, USA) for 60 min. rhGrem1 and BMP proteins were co-incubated in complete medium at 37 o C for 15 min prior to adding to cells.To assess the effect of cell associated Grem1, HK-2 cells were pre-treated with HK-2 complete medium supplemented with vehicle (PBS), 25 ng/ml Grem1 (for BMP-2) or 200 ng/ml Grem1 (for BMP-4) for 60 min. The medium was removed and replaced with fresh HK-2 complete medium supplemented with vehicle (4 mM HCl), 5 ng/ml BMP-2 or 5 ng/ml BMP-2 plus 25 ng/ml rhGrem1. A similar approach was employed for BMP-4, with 5 ng/ml BMP-4 and 200 ng/ml rhGrem1 utilized.

Church R.H., Krishnakumar A., Urbanek A., Geschwindner S., Meneely J., Bianchi A., Basta B., Monaghan S., Elliot C., Strömstedt M., Ferguson N., Martin F, & Brazil D.P. (2015). Gremlin1 preferentially binds to bone morphogenetic protein-2 (BMP-2) and BMP-4 over BMP-7. The Biochemical journal, 466(1).

+ Open protocol

+ Expand

Cell Cycle Analysis of NPC Fate

Check if the same lab product or an alternative is used in the 5 most similar protocols

NPCs were cultured in monolayer with rh-BMP7 and/or rh-FGF9 (50 and 100 ng ml⁻¹, R&D systems) in KSFM (Thermo Fisher Scientific) for 24 h. Cells were fixed in 4% PFA and blocked in 1X PBS containing serum of the secondary antibody species, following which they were incubated in primary antibodies to CCNE1 and PCNA (1:200, Santacruz). Alexa-Fluor-488 (CCNE1) and Alexa-Fluor-568 (PCNA) secondary antibodies were used to visualize the staining. Images (5–8) were taken per well for each condition with a minimum of three biological replicates and three independent experiments (n=3). Pooled images were analysed by Image-J and number of cells positive for G1 (CCNE1+), G1-S (CCNE1+/PCNA+) and S (PCNA+) phases were counted and divided by the total number of DAPI+ nuclei to determine the percentage of cells representing G1, G1-S or S-phase. Data are represented as percentage of G1 or S-phase cells in each condition.

Muthukrishnan S.D., Yang X., Friesel R, & Oxburgh L. (2015). Concurrent BMP7 and FGF9 signalling governs AP-1 function to promote self-renewal of nephron progenitor cells. Nature Communications, 6, 10027.

+ Open protocol

+ Expand

BMP7, FGF9, TAK1i, and JNKi Effects on Protein Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols

Isolated E17.5 NPCs were treated with rh-BMP7 or rh-FGF9 (50 and 100 ng ml⁻¹, R&D systems), TAK1i (0.5 μM, Analyticon Discovery) JNKi (10 μM, Calbiochem) in KSFM (Thermo Fisher Scientific). Total protein was extracted as previously described7 (link). Antibodies used were: anti-pTAK1 (Thr184/187), anti-pJNK (Thr183/Tyr185), anti-pJUN (S73), anti-pFOS (S32) (1:1,000, Cell signaling), anti-β-tubulin (1:5,000, Santa Cruz). Protein levels were quantified using FIJI/Image-J software by measuring the integrated density of the indicated proteins normalized to β-Tubulin, the loading control. Average values from two independent experiments (n=2) are indicated in the graph.

+ Open protocol

+ Expand

Neural Progenitor Cell Proliferation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

E17.5 NPCs were cultured in monolayer with rh-BMP7 and rh-FGF9 (50 and 100 ng ml⁻¹, R&D systems) in KSFM. Cultures were incubated with 20 μM EdU (Click-iT EdU Alexa-Fluor 488 Imaging Kit, Life Technologies) 4 h after growth factor stimulation and pulse-chased for 20 h. Fixation, permeabilization and Click-iT reaction was performed according to manufacturer's instructions. Cultures were incubated with anti-pHH3 (1:100, Cell Signaling) antibody for 1 h and Alexa-Fluor-568 secondary antibody was used to visualize the staining. Nuclei were stained with Hoechst 33342 (Life Technologies). Images (5–10)were taken per well for each condition with a minimum of three biological replicates from two independent experiments (n=2). Pooled images were analysed by Image-J and number of EdU+ (S-phase) and/or pHH3+ (Mitosis or M-phase) nuclei were counted and divided by the total number of nuclei to determine the percentage of cells in S- and M-phases. Data are represented as percentage of S- or M- phase cells in each condition.

+ Open protocol

+ Expand

Preparation of Recombinant BMPs

Check if the same lab product or an alternative is used in the 5 most similar protocols

Commercially available rhBMP-2, rhBMP-4, and rhBMP-7 were purchased from R&D Systems (Minneapolis, MN, USA) and dissolved in sterilized 4 mM HCl (Kanto Chemical Co., Inc., Tokyo, Japan) containing 0.2% BSA (Invitrogen, Carlsbad, CA, USA) before use.

Hayashi T., Asakura M., Kawase M., Matsubara M., Uematsu Y., Mieki A, & Kawai T. (2022). Bone Tissue Engineering in Rat Calvarial Defects Using Induced Bone-like Tissue by rhBMPs from Immature Muscular Tissues In Vitro. International Journal of Molecular Sciences, 23(13), 6927.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!