Human recombinant rank ligand

Manufactured by Thermo Fisher Scientific

Sourced in United States

Human recombinant RANK Ligand is a protein that functions as a ligand for the RANK (receptor activator of nuclear factor kappa-B) receptor. It plays a crucial role in the regulation of bone metabolism, immune system function, and other physiological processes. This product is intended for research use only and its specific applications should be evaluated by the user.

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Lab products found in correlation

Human m csf, by Thermo Fisher Scientific (2 mentions) Dexamethasone (dex), by Merck Group (2 mentions) Human xl oncology array kit, by R&D Systems (1 mentions) Ccl20, by R&D Systems (1 mentions) Quantikine elisa kit, by R&D Systems (1 mentions) Retinoic acid, by Bio-Techne (1 mentions) Human il 1β, by R&D Systems (1 mentions) Mm11253, by Bio-Techne (1 mentions) Bms493, by Bio-Techne (1 mentions) Er50891, by Bio-Techne (1 mentions) Le135, by Bio-Techne (1 mentions) Human wnt3a, by R&D Systems (1 mentions) Recombinant human ifn γ, by R&D Systems (1 mentions) Tamoxifen, by Merck Group (1 mentions) Retinol, by Merck Group (1 mentions) 5 n ethyl n isopropyl amiloride eipa, by Merck Group (1 mentions) Raw 264.7 cells, by American Type Culture Collection (1 mentions) Raw264, by Thermo Fisher Scientific (1 mentions) Z devd fmk, by R&D Systems (1 mentions) Mm 1s, by American Type Culture Collection (1 mentions)

5 protocols using human recombinant rank ligand

Epithelial-Mesenchymal Transition Markers

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Recombinant human RANK ligand and recombinant human MIP-3α/CCL20 were purchased from PeproTech (Rocky Hill, NJ, USA). Rabbit monoclonal antibodies (mAb) against E-cadherin and Vimentin were obtained from Cell Signaling Technology (Danvers, MA, USA). Rabbit polyclonal antibody against N-cadherin, Snail and Twist was from Abcam. Mouse mAb against FLAG was purchased from Cell Signaling Technology. Mouse mAb against RANK and goat polyclonal antibody against RANKL were from SANTA CRUZ (Delaware Avenue, CA, USA). Goat polyclonal antibody CCL20, Human XL Oncology Array Kit and Quantikine ELISA Kit were all purchased from R & D Systems (USA).

Liu Y., Wang J., Ni T., Wang L., Wang Y, & Sun X. (2016). CCL20 mediates RANK/RANKL-induced epithelial-mesenchymal transition in endometrial cancer cells. Oncotarget, 7(18), 25328-25339.

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Signaling Pathways in Cell Differentiation

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Recombinant human IFN-γ was purchased from the R&D systems. Recombinant human LT-α2β1 (#679-TX-010), murine LT-α2β1 (#1008-LY-010), human IL-1β, human IL-22, human Wnt-3a, and anti-LT-β receptor antibody (#AF629) were also purchased from the R&D systems. Recombinant human RANK ligand was purchased from PeproTech (#310-01). Retinoic acid (#695) and RAR inhibitors ER50891, LE135, MM11253, BMS-493 were purchased from Tocris. Retinol and tamoxifen were acquired from Sigma-Aldrich and RARα agonist AM580 was purchased from Wako (Alpha Laboratories). Red fluorescence labeled carboxylate-modified polystyrene latex beads with 0.5 μm diameter were purchased from Sigma-Aldrich. Antibody used for RelB chromatin immunoprecipitation (sc-226) was purchased from Santa Cruz Biotechnology.

Ding S., Song Y., Brulois K.F., Pan J., Co J.Y., Ren L., Feng N., Yasukawa L.L., Sánchez-Tacuba L., Wosen J.E., Mellins E.D., Monack D.M., Amieva M.R., Kuo C.J., Butcher E.C, & Greenberg H.B. (2020). Retinoic Acid and Lymphotoxin Signaling Promote Differentiation of Human Intestinal M Cells. Gastroenterology, 159(1), 214-226.e1.

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Differentiation of Murine Macrophages

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U266, MM1s and OPM2 cell lines were purchased from ATCC^® (LGC Standards S.r.l.Sesto San Giovanni, MI, Italy) and grown in RPMI-1640 (Gibco, Life Technologies, USA) supplemented with 10% Fetal Bovine Serum (FBS, Lonza Group, Basel, Switzerland).
Murine macrophage Raw264.7 cells were purchased from ATCC^® and cultured in Dulbecco's modified Eagle's medium (DMEM), supplemented with 10% FBS. To induce differentiation, cells were treated with 25ng/ml of human recombinant RANK Ligand (Gibco, Life Technologies, USA) for 6 days in DMEM, supplemented with 10% FBS, previously ultracentrifugated (Raw264.7 cells OC medium). Alternately, cells were treated for 6 days with 25μg/ml of MM cell-derived exosomes, in DMEM, supplemented with 10% of ultracentrifugated FBS (see below). 5-(N-Ethyl-N-isopropyl) amiloride (EIPA) was purchased from Sigma Aldrich-Italy. Caspase-3 inhibitor Z-DEVD-FMK was purchased from R&D systems.

Raimondi L., De Luca A., Amodio N., Manno M., Raccosta S., Taverna S., Bellavia D., Naselli F., Fontana S., Schillaci O., Giardino R., Fini M., Tassone P., Santoro A., De Leo G., Giavaresi G, & Alessandro R. (2015). Involvement of multiple myeloma cell-derived exosomes in osteoclast differentiation. Oncotarget, 6(15), 13772-13789.

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Human Osteoclastogenesis Assay Protocol

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PBMCs were cultured at 2.5 × 10⁶ cells/mL α-Minimum Essential Media (MEM) supplemented with 10% FBS previously ultracentrifuged, 25 ng/mL of human recombinant RANK Ligand (Gibco, Life Technologies, Rockford, IL, USA), 25 ng/mL of human M-CSF (Gibco, Thermo Fisher Scientific, Rockford, IL, USA), and 10 nM dexamethasone (Sigma-Aldrich, Milano, Italy) (Human OC medium). After 3 days, the culture were washed with α-MEM medium to remove non adherent cells. The remaining cells were mononucleated, expressed TRAP and were considered committed pre-osteoclast cells. For human osteoclastogenesis assays, human OC medium was added and the cultures were continued for additional 6–10 days, at the end of the period they contained large mature multinucleated OCs. The culture period was 14 days for both TRAP staining assay, bone resorption assay and ELISA assay.

Raimondi L., De Luca A., Fontana S., Amodio N., Costa V., Carina V., Bellavia D., Raimondo S., Siragusa S., Monteleone F., Alessandro R., Fini M, & Giavaresi G. (2020). Multiple Myeloma-Derived Extracellular Vesicles Induce Osteoclastogenesis through the Activation of the XBP1/IRE1α Axis. Cancers, 12(8), 2167.

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Osteoclastogenesis from Human PBMCs

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PBMCs were cultured at 2.5×10⁶cells/ml α-MEM supplemented with 10% FBS previously ultracentrifuged, 25ng/ml of human recombinant RANK Ligand (Gibco, Life Technologies, USA), 25ng/ml of human M-CSF (Gibco, Life Technologies, USA), and 10nM dexamethasone (Sigma-Aldrich Italy) (Human OC medium). After 2-4 days, the culture were washed with α-MEM medium to remove non adherent cells. The remaining cells were mononucleated, expressed TRAP and were considered committed pre-osteoclast cells. For human osteoclastogenesis assays, OC medium was added and the cultures were continued for additional 6-10 days, at the end of the period they contained large mature multinucleated OCs. The culture period was 16 days for both TRAP staining assay, bone resorption assay and qRT-PCR analysis.

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