Recombinant bmp7

Manufactured by R&D Systems

Sourced in United States

Recombinant BMP7 is a recombinant human Bone Morphogenetic Protein 7. It is a secreted protein that plays a role in the regulation of bone and cartilage development.

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Lab products found in correlation

Nih 3t3, by Leibniz Institute DSMZ (1 mentions) Epi 2500 impulse generator, by Thermo Fisher Scientific (1 mentions) Pcmv6 xl4, by OriGene (1 mentions) A83 01, by Bio-Techne (1 mentions) Megm complete media, by Lonza (1 mentions) Ham s f12 medium, by Thermo Fisher Scientific (1 mentions) Quikchange site directed mutagenesis kit, by Agilent Technologies (1 mentions) Penicillin streptomycin glutamine (psg), by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Gemini Bio (1 mentions) Rabbit anti gapdh igg, by Abcam (1 mentions) Dmem high glucose medium, by Thermo Fisher Scientific (1 mentions) Rabbit anti gfp igg, by Thermo Fisher Scientific (1 mentions) Multiscan jx, by Thermo Fisher Scientific (1 mentions)

4 protocols using recombinant bmp7

Modulating gene expression in DLBCL and HL cell lines

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DLBCL cell lines, HL cell line L-540, and NIH-3T3 cells were obtained from the DSMZ (Braunschweig, Germany) and cultivated as described elsewhere [33 ]. To modify gene expression levels we used gene specific siRNA oligonucleotides in comparison to AllStars negative Control siRNA (siControl) which were obtained from Qiagen (Hilden, Germany). Gene expression constructs for HHEX, HLX, MSX1 and PCGF5 were cloned in vector pCMV6-XL4 and obtained from Origene (Wiesbaden, Germany). SiRNAs (80 pmol) and expression constructs/vector controls (2 μg) were transfected into 1x10⁶ cells by electroporation using the EPI-2500 impulse generator (Fischer, Heidelberg, Germany) at 350 V for 10 ms. Electroporated cells were harvested after 20 h cultivation. Cells were treated for 16 h with 20 ng/ml recombinant BMP7 (R & D Systems, Abingdon, UK), with BMP receptor inhibitor dorsomorphin (DM) (Calbiochem, Darmstadt, Germany) dissolved in dimethyl sulfoxide (DMSO) at a final concentration of 5 μM.

Nagel S., MacLeod R.A., Meyer C., Kaufmann M, & Drexler H.G. (2018). NKL homeobox gene activities in B-cell development and lymphomas. PLoS ONE, 13(10), e0205537.

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Immortalized Mammary Epithelial Cell Culture

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Immortalized mammary epithelial cells (IMECs) and IMEC-sh-p53 cells were derived as previously described (14 (link), 15 (link)). IMECs were cultured in MEGM complete media (Lonza CC-3051) with 50 μg/ml puromycin, Bovine Pituitary Extract (BPE), and 100 units/ml Penicillin/Streptomycin. Treatments with TGFβ1 and A83-01 were performed without BPE and with the addition of 0.1%BSA. H1299 cells were cultured according to ATCC guidelines. Cells were serum starved for 3 hours prior to all TGFβ treatments. Recombinant TGFβ1 (Miltenyi) was used at 500pM. A83-01 (Tocris) was used at 2uM for 1 hr prior to TGFβ1 treatment, or as indicated. LDN193189 (Stemgent Technologies) was used at 1uM, unless otherwise indicated. Recombinant BMP7 (R&D systems) was used at 50 ng/ml.

Balboni A.L., Cherukuri P., Ung M., DeCastro A.J., Cheng C, & DiRenzo J. (2015). p53 and ΔNp63α Co-regulate the Transcriptional and Cellular Response to TGFβ and BMP Signals. Molecular cancer research : MCR, 13(4), 732-742.

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BMP7 Signaling Analysis Protocol

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Recombinant BMP7 was purchased from R&D Systems (Minneapolis, MN, USA). Antibodies: mouse anti-ERM (Ezrin/Radixin/Moesin) IgM (13H9; Goslin et al., 1989 (link)); rabbit anti-GFP IgG (Invitrogen); rabbit anti-ActRII IgG (H65) and HRP-conjugated secondary antibodies (Santa Cruz Biotechnology); rabbit anti-phospho-Smad1/5/8 IgG (pSmad), rabbit anti-Smad1 IgG (tSmad), rabbit anti-phopho-Akt IgG (pAkt) and rabbit anti-Akt IgG (tAkt) (Cell Signaling Technology); rabbit anti-GAPDH IgG (Abcam); Cy3- and Cy2-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories). The dilutions for each antibody used in the study are listed in Table S1. Cell culture reagents: Ham's F12 medium, DMEM high-glucose medium, 100× Penicillin/Streptomycin/Glutamine (PSG) (Invitrogen), FBS (Gemini BioProducts, West Sacramento, CA, USA). The expression construct for flag-tagged, mouse ActRIIA.pcDNA3 was generously provided by Dr K. Miyazono (The JFCR Cancer Institute, Japan). Site-directed mutagenesis of ActRIIA V3 at Lys⁷⁶ and of ActRIIB at Glu⁷⁵ was carried out (see Fig. 4) using the QuikChange Site-Directed Mutagenesis Kit (Agilent Technologies, Santa Clara, CA, USA).

Perron J.C., Rodrigues A.A., Surubholta N, & Dodd J. (2019). Chemotropic signaling by BMP7 requires selective interaction at a key residue in ActRIIA. Biology Open, 8(7), bio042283.

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BMP-7 Dose-Dependent Cell Proliferation

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Cells were seeded at a density of 5000/ml in 100-ml medium, and recombinant BMP-7 (R&D Systems, Minneapolis, MN, USA) was added at concentrations of 0, 10, 50, or 100 ng/ml (n ¼ 5 for each test) as previously reported. 28, 29 Cells were incubated at 37 C for 24 h, and proliferation was analyzed according to the manufacturer's instructions (Promega Corporation, Madison, WI, USA). Twenty microliters of MTS reagent was added directly to the wells and incubated for 3 h. Absorbance was measured at 492 nm on a Multiscan JX microplate reader (Thermo Labsystems, Waltham, MA, USA). The results are represented as the percentage of relative fluorescence units compared with the control.

Kokubu N., Tsujii M., Akeda K., Iino T, & Sudo A. (2018). BMP-7/Smad expression in dedifferentiated Schwann cells during axonal regeneration and upregulation of endogenous BMP-7 following administration of PTH (1-34). Journal of orthopaedic surgery (Hong Kong), 26(3).

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