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Recombinant opg

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Recombinant OPG is a recombinant protein manufactured by R&D Systems. It is a member of the tumor necrosis factor receptor superfamily and functions as a regulator of bone remodeling.

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Lab products found in correlation

Purefection reagent, by System Biosciences (3 mentions) Anti cleaved caspase 3, by Cell Signaling Technology (1 mentions) Recombinant trail, by R&D Systems (1 mentions) Halt protease and phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions) Anti vinculin, by Cell Signaling Technology (1 mentions) Anti caspase 3, by Cell Signaling Technology (1 mentions) Clarity western ecl substrate, by Bio-Rad (1 mentions) D glucose, by Fujifilm (1 mentions) Bca protein assay reagent, by Thermo Fisher Scientific (1 mentions) Soluble recombinant rankl, by R&D Systems (1 mentions) Glucose, by Cosmo Bio (1 mentions) Recombinant m csf, by Thermo Fisher Scientific (1 mentions) Anti β actin antibody ac 74, by Merck Group (1 mentions) Recombinant mouse s100a4, by ProSpec (1 mentions) Anti s100a4 antibody, by Abcam (1 mentions) Anti β catenin antibody, by Thermo Fisher Scientific (1 mentions) Rankl, by Thermo Fisher Scientific (1 mentions) Leukocyte acid phosphatase kit, by Merck Group (1 mentions) Dentin slices, by Immunodiagnostic Systems (1 mentions) Penicillin streptomycin, by American Type Culture Collection (1 mentions)

4 protocols using recombinant opg

1

OPG Modulates TRAIL-Induced Apoptosis

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MDA231-LM2 cells, treated with incremental dosages of recombinant OPG (10–200 ng ml–1, R&D Systems) in the presence of 50 ng ml–1 recombinant TRAIL (R&D Systems), were lysed in RIPA buffer with 1× HALT protease and phosphatase inhibitor cocktail (Thermo Fisher Scientific). Immunoblots were performed as previously reported51 . Primary antibodies used were as follows: anti-cleaved caspase 3 (1:500), anti-caspase 3 (1:1,000) and anti-vinculin (1:1,000), all from Cell Signaling. Incubation with horseradish peroxidase-conjugated IgG (1:10,000, Leica), followed by Clarity Western ECL Substrate (Bio-Rad) and exposure to X-ray films (Fuji-film) was used to develop the signal.
Hongu T., Pein M., Insua-Rodríguez J., Gutjahr E., Mattavelli G., Meier J., Decker K., Descot A., Bozza M., Harbottle R., Trumpp A., Sinn H.P., Riedel A, & Oskarsson T. (2022). Perivascular tenascin C triggers sequential activation of macrophages and endothelial cells to generate a pro-metastatic vascular niche in the lungs. Nature Cancer, 3(4), 486-504.
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2

Insulin Secretion Assay in MIN6 Cells

Cited 1 time
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A method to assay insulin secretion in vitro using MIN6 cells [53 (link)] has been described [54 (link)]. Briefly, MIN6 cells were maintained in Dulbecco’s modified Eagle’s medium containing 4.5 g/L D-glucose (Wako), 10% fetal bovine serum, 0.1 mM 2-mercaptoethanol, 100 units/ml penicillin, and 0.05 mg/ml streptomycin in humidified 5% CO2 at 37°C. Cells were replated and cultured for 3 days, and then treated with 10 μg/ml LPS, 100 ng/ml recombinant soluble RANKL (R&D systems), or 100 ng/ml recombinant OPG (R&D systems) as indicated. After additional culture for 24 hr, cells were starved in HEPES-balanced Krebs-Ringer bicarbonate (HKRB) buffer with 3 mM glucose (Cosmobio) for 30 min and then incubated 1 hr in HKRB buffer containing 3, 9.8, or 20 mM D-glucose. Culture supernatants were collected for insulin measurement. Cell lysates were prepared with RIPA buffer to measure protein using the BCA Protein Assay Reagent (Thermo Fisher Scientific).
Kuroda Y., Maruyama K., Fujii H., Sugawara I., Ko S.B., Yasuda H., Matsui H, & Matsuo K. (2016). Osteoprotegerin Regulates Pancreatic β-Cell Homeostasis upon Microbial Invasion. PLoS ONE, 11(1), e0146544.
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3

Osteoclastogenesis Signaling Pathway

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Recombinant M-CSF and RANKL were purchased from PeproTech. Recombinant mouse S100A4 was purchased from Prospec. Recombinant OPG was purchased from R&D Systems. Antibodies against vimentin and c-Fos were purchased from Santa Cruz. Antibodies against N-cadherin and snail2 were obtained from Cell Signaling Technology. Antibodies for RAGE (DD/A11) were from Millipore. Anti-S100A4 antibody was purchased from Abcam. Anti-β-catenin antibody was purchased from Invitrogen. Antibodies for NFATc1 (7A6), E-cadherin, and hHLA were purchased from BD Pharmingen. Anti-β-actin antibody (AC-74) and the leukocyte acid phosphatase kit (for TRAP staining) were purchased from Sigma-Aldrich. Lipofectamine for siRNA transfection was purchased from Life Technologies. siRNA oligonucleotides and shRNA lentiviral particles were purchased from Santa Cruz. Dentin slices were purchased from Immunodiagnostic Systems.
Kim B., Jung S., Kim H., Kwon J.O., Song M.K., Kim M.K., Kim H.J, & Kim H.H. (2021). The role of S100A4 for bone metastasis in prostate cancer cells. BMC Cancer, 21, 137.
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4

Establishing Osteolytic Prostate Cancer Cell Line PC3 for Research

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The human osteolytic prostate cancer cell line PC3, expressing firefly luciferase, (a kind gift from Dr. Kenneth J. Pienta, University of Michigan, Ann Arbor, Michigan), was maintained in RPMI-1640 medium (Mediatech Inc. Hendron, VA) supplemented with 10% fetal bovine serum (Mediatech Inc.) and penicillin/streptomycin (Mediatech Inc). The murine macrophage cell line, RAW-264.7 was maintained in DMEM supplemented with 10% FBS, 4 mM L-glutamine, 1% antibiotics and 10% macrophage colony-stimulating factor (M-CSF, a kind gift from Dr. Xu Feng, The University of Alabama at Birmingham). The human melanoma cell line MDA-MB-435 was maintained in 50% DMEM, 50% DMEM F12, 1% penicillin/streptomycin, 10% FBS, and non-essential amino acids. HEK-293 cells were purchased from ATCC and maintained in DMEM supplemented with 10% new born calf serum and 1% penicillin-streptomycin. The proliferation index of PC3 cells was determined by CellTiter 96® AQueous One Solution Cell Proliferation Assay Kit (Promega Corporation, Madison, WI) as recommended by the manufacturer. Recombinant OPG was purchased from R&D Systems and recombinant TRAIL was purchased from Millipore. Purified RANKL was a kind gift from Dr. Xu Feng, UAB. Transfections were performed using Purefection reagent (System Bioscience, Mountain View, CA).
Higgs J.T., Jarboe J.S., Lee J.H., Chanda D., Lee C.M., Deivanayagam C, & Ponnazhagan S. (2015). Variants of Osteoprotegerin Lacking TRAIL Binding for Therapeutic Bone Remodeling in Osteolytic Malignancies. Molecular cancer research : MCR, 13(5), 819-827.
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