Rhrankl

Manufactured by Thermo Fisher Scientific

Sourced in Germany

RhRANKL is a recombinant protein produced in a mammalian expression system. It is a member of the tumor necrosis factor (TNF) ligand family and plays a key role in the regulation of bone remodeling and the immune system.

Automatically generated - may contain errors

Lab products found in correlation

Rhm csf, by Thermo Fisher Scientific (6 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Penicillin streptomycin, by Merck Group (2 mentions) L glutamine, by Merck Group (2 mentions) Rhm csf, by R&D Systems (2 mentions) Primary and secondary antibodies, by Cell Signaling Technology (1 mentions) Ammonium bicarbonate, by Merck Group (1 mentions) Sodium deoxycholate, by Merck Group (1 mentions) Dithiothreitol (dtt), by Thermo Fisher Scientific (1 mentions) Iodoacetamide, by Thermo Fisher Scientific (1 mentions) Methanol, by Avantor (1 mentions) Rapigest sf surfactant, by Waters Corporation (1 mentions) α mem, by Merck Group (1 mentions) Ficoll, by Merck Group (1 mentions) Trypsin gold, by Promega (1 mentions) Trypsin lys c mix, by Promega (1 mentions) Trypsin edta, by Thermo Fisher Scientific (1 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions) Acetonitrile, by Merck Group (1 mentions) Formic acid, by Merck Group (1 mentions)

9 protocols using rhrankl

Osteoclastogenesis Assay with DARA and ATRA

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

OCs were obtained either from total BM MNCs of MM patients, or purified CD14⁺ cells from PBMCs of 3 HDs. Cells were seeded in αMEM with 10% FBS, rhM-CSF 25 ng/ml and rhRANKL 60 ng/ml (Peprotech; Rocky Hill, NJ) in the presence of DARA (10–25 μg/ml) or isotype control IgG added to culture medium on day 0 or after 10 days, with or without ATRA (0.1–20 nM) or vehicle (DMSO) for 21 days, replacing half medium every 2–3 days. BM MNCs were also incubated in the presence or absence of the CM (dilution ratio with αMEM with 10% FBS, rhM-CSF 25 ng/ml and rhRANKL 60 ng/ml, 1:3) of JJN3 and RPMI-8226 (5 × 10⁵ cells/ml) previously pre-treated with DARA (10-25 μg/ml) or isotype control IgG for 48 h, and cultured for 21 days replacing half medium every 2-3 days. Each condition was performed at least in triplicate. For dose-finding experiment, BM MNCs were cultured in αMEM with 10% FBS, rhM-CSF 25 ng/ml and rhRANKL 60 ng/ml (Peprotech; Rocky Hill, NJ) in presence of ATRA (0.01 nM–200 nM), or vehicle (DMSO) for 21 days, replacing half medium every 2–3 days.
MVs isolation, tartrate resistant acid phosphatase (TRAP) and Osteoclastogenesis resorption assays. Methods are detailed in the Supplementary Methods.

Costa F., Toscani D., Chillemi A., Quarona V., Bolzoni M., Marchica V., Vescovini R., Mancini C., Martella E., Campanini N., Schifano C., Bonomini S., Accardi F., Horenstein A.L., Aversa F., Malavasi F, & Giuliani N. (2017). Expression of CD38 in myeloma bone niche: A rational basis for the use of anti-CD38 immunotherapy to inhibit osteoclast formation. Oncotarget, 8(34), 56598-56611.

+ Open protocol

+ Expand

Osteoclastogenesis Induction Protocol

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

Human recombinant (rh)M-CSF and rhRANKL were purchased from Peprotech (Hamburg, Germany); cell culture medium and supplements were purchased from PAA (Cölbe, Germany); primary and secondary antibodies were obtained from Cell Signaling (Beverly, USA); chemicals were purchased from Sigma (Munich, Germany).

Ehnert S., Falldorf K., Fentz A.K., Ziegler P., Schröter S., Freude T., Ochs B.G., Stacke C., Ronniger M., Sachtleben J, & Nussler A.K. (2015). Primary human osteoblasts with reduced alkaline phosphatase and matrix mineralization baseline capacity are responsive to extremely low frequency pulsed electromagnetic field exposure — Clinical implication possible. Bone Reports, 3, 48-56.

+ Open protocol

+ Expand

Proteomic Analysis of Macrophage Differentiation

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

Ficoll, MEM α, L-glutamine, and Penicillin–Streptomycin were obtained from Sigma (St. Louis, MO, USA). FBS, PBS, and trypsin-EDTA solutions were purchased from Gibco Laboratories (Gaithersburg, MD, USA). EasySep was obtained from StemCell Technologies (Vancouver, Canada), rh M-CSF, and rh RANKL were purchased from PeproTech (London, UK).
Acetonitrile, water, formic acid, ammonium-bicarbonate, and sodium deoxycholate were obtained from Merck (Darmstadt, Germany). Methanol was purchased from VWR International (Debrecen, Hungary). Dithiothreitol, iodoacetamide, and TFA were purchased from Thermo Scientific (Waltham, MA, USA), and RapiGest SF surfactant was obtained from Waters (Milford, MA, USA). Trypsin/Lys-C Mix and Trypsin Gold were purchased from Promega Corporation (Madison, WI, USA). All chemicals, reagents, and solvents were HPLC-MS grade.

Kovács O.T., Tóth E., Ozohanics O., Soltész-Katona E., Marton N., Buzás E.I., Hunyady L., Drahos L., Turu G, & Nagy G. (2022). Proteomic Changes of Osteoclast Differentiation in Rheumatoid and Psoriatic Arthritis Reveal Functional Differences. Frontiers in Immunology, 13, 892970.

+ Open protocol

+ Expand

Recombinant Cytokine-Stimulated Cell Culture

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

rhM-CSF and rhRANKL were purchased from Peprotech (Hamburg, Germany). Chemicals, cell culture medium, and supplements were purchased from Sigma-Aldrich (Munich, Germany).

Ehnert S., van Griensven M., Unger M., Scheffler H., Falldorf K., Fentz A.K., Seeliger C., Schröter S., Nussler A.K, & Balmayor E.R. (2018). Co-Culture with Human Osteoblasts and Exposure to Extremely Low Frequency Pulsed Electromagnetic Fields Improve Osteogenic Differentiation of Human Adipose-Derived Mesenchymal Stem Cells. International Journal of Molecular Sciences, 19(4), 994.

+ Open protocol

+ Expand

Osteoclastogenesis from Murine Bone Marrow

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

Mouse bone marrow cells derived from less than 8 weeks old mice were plated in 96-well plates in osteoclastic medium containing a-MEM supplemented with 10% FBS, 100 U/mL of penicillin, 100 mg/mL of streptomycin, 25 ng/mL of recombinant human macrophage colony-stimulating factor (rhM-CSF; R&D Systems, Minneapolis, MN, USA), and 25 ng/ mL of recombinant human receptor activator of NF-kB (rhRANKL; Pepro-Tech, Rocky Hill, NJ, USA). The presence of osteoclastic cells was verified by staining for tartrate-resistant acid phosphatase (TRAcP) performed 5-7 days after culture, and TRAcP multinucleated (≥ 3) cells were scored as osteoclastic cells.

Chen L., Shi K., Ditzel N., Qiu W., Figeac F., Nielsen L.H., Tencerova M., Kowal J.M., Ding M., Andreasen C.M., Andersen T.L, & Kassem M. (2023). KIAA1199 deficiency enhances skeletal stem cell differentiation to osteoblasts and promotes bone regeneration. Nature Communications, 14, 2016.

+ Open protocol

+ Expand

Isolation and Differentiation of Human Monocytes

Cited 1 time

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

Human blood samples were collected into vacutainer EDTA tubes (Greiner Bio-One, Mosonmagyarovar, Hungary). PBMCs (peripheral blood mononuclear cells) were isolated via a Ficoll density gradient (Sigma, St. Louis, MO, USA). CD14+ cells were isolated from PBMCs using a positive magnetic selection method (EasySep, StemCell Technologies, Vancouver, Canada) (29 (link)). After isolation, a third of the monocytes were immediately frozen in liquid nitrogen. Two-thirds of monocytes were cultured (1x10⁵ monocytes per well) in MEM α (Minimum Essential Medium α) (Sigma, St. Louis, MO, USA) with 10% FBS (Gibco Laboratories, Gaithersburg, MD, USA), 1% L-glutamine (Sigma, St. Louis, MO, USA), and 1% Penicillin–Streptomycin (Sigma, St. Louis, MO, USA) at 37°C and 5% CO₂ in a humidified atmosphere. Monocytes were incubated with 50 ng/mL rh M-CSF (macrophage colony-stimulating factor) for 24 h (PeproTech, London, UK). Thereafter the samples were stimulated with 50 ng/mL of both rh M-CSF and rh RANKL (receptor activator of nuclear factor κB ligand) (PeproTech, London, UK). From then the media was replaced every 3-4 days.

+ Open protocol

+ Expand

Isolation and Differentiation of Monocyte-Derived Cells

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

CD14⁺ monocytes were isolated from fresh PBMCs with CD14 MicroBeads (Miltenyi Biotec, 130-050-201) according to the manufacturer’s instructions. For the induction of monocyte-derived macrophages (MDMs), CD14⁺ monocytes were cultured in RPMI 1640 supplemented with 10% FCS, 1% penicillin/streptomycin and 50 ng/ml recombinant human (rh)M-CSF (R&D Systems, 216-MC) for 7 days, and were then allowed to differentiate for seven days in the presence of 50 ng/ml rhM-CSF and 50 ng/ml rhIL-4 (R&D Systems, 204-IL). Monocyte-derived dendritic cells (MDDCs) were induced by culturing CD14⁺ monocytes for 7 days in RPMI 1640 supplemented with 10% FCS, 1% penicillin/streptomycin, 50 ng/ml rhGM-CSF (R&D Systems, 215-GM) and 20 ng/ml rhIL-13 (R&D Systems, 213-ILB). For osteoclast differentiation, CD14⁺ cells were cultured in α-MEM (Thermo Fisher Scientific, 12571063) containing 10% FCS, 1% penicillin/streptomycin, 30 ng/ml rhM-CSF and 10 ng/ml rhRANKL (Peprotech, 310-01) for 7 days. The medium was replaced every three days.

Bohlen J., Zhou Q., Philippot Q., Ogishi M., Rinchai D., Nieminen T., Seyedpour S., Parvaneh N., Rezaei N., Yazdanpanah N., Momenilandi M., Conil C., Neehus A.L., Schmidt C., Franco C.A., Le Voyer T., Khan T., Yang R., Puchan J., Erazo L., Roiuk M., Vatovec T., Janda Z., Bagaric I., Materna M., Gervais A., Li H., Rosain J., Peel J., Seeleuthner Y., Han J.E., L’Honneur A.S., Moncada-Vélez M., Martin-Fernandez M., Horesh M.E., Kochetkov T., Schmidt M., AlShehri M.A., Salo E., Harri S., ElGhazali G., Yatim A., Soudée C., Sallusto F., Ensser A., Marr N., Zhang P., Bogunovic D., Cobat A., Shahrooei M., Béziat V., Abel L., Wang X., Boisson-Dupuis S., Teleman A.A., Bustamante J., Zhang Q, & Casanova J.L. (2023). Human MCTS1-dependent translation of JAK2 is essential for IFN-γ immunity to mycobacteria. Cell, 186(23), 5114-5134.e27.

+ Open protocol

+ Expand

Osteoclast Formation Assay

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

For osteoclast formation, 2.5 × 10³ cells/cm² were seeded into the wells of a 24-well plate. After pre-culture, the medium was changed to media containing 100 ng/ml recombinant human receptor activator of nuclear factor κ-B ligand (rhRANKL; PeproTech, Rocky Hill, NJ) with or without soy isoflavones and carotenoids. The addition of each reagent solvent to the cultured medium was used as a control. After 4 days, the cells were fixed and stained for TRAP activity.^{(10 )} Briefly, cells were washed three times with Dulbecco’s phosphate-buffered saline (PBS) and fixed in a solution of ethanol-acetone (1:1). After drying, the cells were stained in 0.1 M sodium acetate buffer (pH 5.0) containing 50 mM sodium tartrate, 0.1 mg/ml naphthol AS-MX phosphate (Sigma-Aldrich), and 0.5 mg/ml fast red violet LB salt (Sigma-Aldrich) at 37°C. TRAP-positive cells containing three or more nuclei were counted as TRAP-positive multinucleated osteoclast-like cells (MNCs).

Tadaishi M., Nishide Y., Tousen Y., Kruger M.C, & Ishimi Y. (2014). Cooperative effects of soy isoflavones and carotenoids on osteoclast formation. Journal of Clinical Biochemistry and Nutrition, 54(2), 109-115.

+ Open protocol

+ Expand

Inhibitory Effect of γδ T Cells on Osteoclast Differentiation

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

γδ T cells and autologous iDCs from the same healthy volunteer were co-cultured at different ratios in Transwell inserts. The γδ T cells were added in the upper and the iDCs were added in the lower compartment of the Transwells, with the medium containing 10% FBS, 100 ng/ml rh-RANKL (PeproTech, Inc.) and 25 ng/ml rh-M-CSF (PeproTech, Inc.). To further confirm if the stage of OC differentiation was important for the inhibitory effect of γδ T cells, γδ T cells were co-cultured with iDCs at the ratio of 1:1 and iDCs were collected at different time-points of culture. The day 0–1 group, day 3–4 group and the day 0–4 group were treated with γδ T cells during days 0–1, day 3–4 and day 0–4, respectively.

Zhu X., Zeng Z., Qiu D, & Chen J. (2018). Vγ9Vδ2 T cells inhibit immature dendritic cell transdifferentiation into osteoclasts through downregulation of RANK, c-Fos and ATP6V0D2. International Journal of Molecular Medicine, 42(4), 2071-2079.

+ 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!