Srankl

Manufactured by Thermo Fisher Scientific

Sourced in United States, United Kingdom

SRANKL is a recombinant human protein that functions as a cell surface receptor involved in the regulation of bone metabolism and the immune system. It plays a role in the differentiation and activation of osteoclasts, the cells responsible for bone resorption.

Automatically generated - may contain errors

Lab products found in correlation

M csf, by Thermo Fisher Scientific (9 mentions) Lamin b, by Abcam (3 mentions) Tumor necrosis factor (tnf), by Thermo Fisher Scientific (3 mentions) Nfatc1, by BD (3 mentions) Tnf α, by Thermo Fisher Scientific (3 mentions) α mem, by Merck Group (2 mentions) Srankl, by Merck Group (2 mentions) Phospho erk, by Cell Signaling Technology (2 mentions) Mtt assay kit, by Roche (2 mentions) Blimp1, by Cell Signaling Technology (2 mentions) I bet, by GlaxoSmithKline (2 mentions) C myc and p38, by Santa Cruz Biotechnology (2 mentions) Macrophage colony stimulating factor m csf, by Thermo Fisher Scientific (2 mentions) Dexamethasone (dex), by Merck Group (2 mentions) Tgf β, by R&D Systems (2 mentions) Trap alp staining kit, by Fujifilm (1 mentions) 6 well plate, by BD (1 mentions) 24 well plate, by BD (1 mentions) Dmi1 microscope, by Leica (1 mentions) Penicillin streptomycin, by Lonza (1 mentions)

Spelling variants of this product

Recombinant murine sRANKL (7 citations) Recombinant murine soluble RANKL (sRANKL) (4 citations) Soluble RANKL (sRANKL, (3 citations) Soluble recombinant mouse RANKL (sRANKL) (2 citations)

22 protocols using srankl

Osteoclastogenesis from RAW264 and MSCs

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

RAW264 cells were seeded in 24-well plates (Falcon) at 5 × 10 3 cells/well. On the following day (day 0), RANK Ligand, Soluble, Murine, Recombinant (sRANKL; Pepro Tech, Rocky Hill, CT, USA) was added to α-MEM (Sigma Aldrich) at 100 ng/mL. The medium was replaced with medium containing sRANKL every 2nd day. On day 8, TRAP staining was performed using a TRAP/ALP staining kit (Wako, Tokyo, Japan) to confirm differentiation into osteoclasts, and cells were observed with a DMi1 microscope (Leica, Wetzlar, Germany).
Next, MSCs were seeded into 6-well plates (Falcon) at 1 × 10 4 cells/well, and RAW264 cells were seeded at In other 6-well plates (Falcon), RAW264 cells were cultured alone at 5 × 10 3 cells/well. On the following day (day 0), the culture medium was replaced with medium containing sRANKL (Pepro Tech) (sRANKL was added to both RAW264 only cultures and RAW264/MSC co-cultures), and the medium was replaced with fresh medium every 2nd day.
From day 0 (at the start of sRANKL addition), using the number of actin ring-forming cells with five or more nuclei to represent the number of mature osteoclasts, osteoclast counts among RAW264 cells cultured alone and in RAW264 cell/MSC co-cultures were compared.

Abe T., Sumi K., Kunimatsu R., Oki N., Tsuka Y., Nakajima K., Ando K, & Tanimoto K. (2019). The effect of mesenchymal stem cells on osteoclast precursor cell differentiation. Journal of oral science, 61(1).

+ Open protocol

+ Expand

Molecular Mechanisms of Osteoclastogenesis

Cited 1 time

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

Human and murine M-CSF, sRANKL and TNF were purchased from Peprotech (Rocky Hill, NJ). I-BET was synthesized by GlaxoSmithKline (UK) as previously described^{15 (link),51 (link),52 (link)}. MYC inhibitor 10058-F4 was purchased from Sigma-Aldrich. The MTT assay kit was purchased from Roche, and cell viability was measured following the manufacturer’s instructions. The antibodies (1:1000) used for immunoblotting are as follows: NFATc1(BD Pharmagen; 556602); c-myc and p38 (Santa Cruz Biotechnology; sc-764 and sc535); Lamin B (Abcam; ab16048); phospho-ERK, IκBα, phospho-p38 and Blimp1 (Cell Signaling; 9101, 9242, 9215, 9115).

Park-Min K.H., Lim E., Lee M.J., Park S.H., Giannopoulos E., Yarilina A., van der Meulen M., Zhao B., Smithers N., Witherington J., Lee K., Tak P.P., Prinjha R.K, & Ivashkiv L.B. (2014). Inhibition of Osteoclastogenesis and Inflammatory Bone Resorption by Targeting BET Proteins and Epigenetic Regulation. Nature communications, 5, 5418.

+ Open protocol

+ Expand

Murine Osteoclastogenesis Assay

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

Dulbecco’s modified eagle’s media (DMEM), minimum essential medium alpha medium (α-MEM), D-glucose, and Alizarin red S dye were supplied by Sigma-Aldrich Chemical (St. Louis, MO, USA), as were all other reagents, unless specifically stated otherwise. Fetal bovine serum (FBS), trypsin–EDTA, and penicillin–streptomycin were obtained from BioWhittaker (San Diego, CA, USA). 3-(4, 5-Dimetylthiazol-yl)-diphenyl tetrazolium bromide (MTT) was purchased from DUCHEFA Biochemie (Haarlem, Netherlands). The recombinant murine sRANKL was purchased from PeproTech (Rocky Hill, NJ, USA). Scopoletin (Sigma-Aldrich Chemical, St. Louis, MO, USA) was dissolved in dimethyl sulfoxide (DMSO) for live culture with cells; the final culture concentration of DMSO was <0.5%.

Lee E.J., Na W., Kang M.K., Kim Y.H., Kim D.Y., Oh H., Kim S.I., Oh S.Y., Park S., Park K, & Kang Y.H. (2021). Hydroxycoumarin Scopoletin Inhibits Bone Loss through Enhancing Induction of Bone Turnover Markers in a Mouse Model of Type 2 Diabetes. Biomedicines, 9(6), 648.

+ Open protocol

+ Expand

Osteoclastogenesis Assay of RAW264.7 Cells

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

Osteoclast differentiation of RAW264.7 co-cultured with MM CM was assessed after 6 days of culture conditions by the detection of TRAP activity, according to the procedure provided by the manufacturer (Acid-phosphatase leukocyte staining kit; Sigma–Aldrich, USA) and observed by optical microscopy. Mature OC are defined as multinucleated TRAP⁺ cells that contain more than 3 nuclei. As control, RAW264.7 cells were incubated with 100 ng/mL sRANKL (PeproTech, USA) and 25 ng/mL macrophage-colony stimulating factor (M-CSF) (PeproTech, USA).

Li T., Qiu D., Chen Q., Yang A., Chen J, & Zeng Z. (2022). NCX1 disturbs calcium homeostasis and promotes RANKL-induced osteoclast differentiation by regulating JNK/c-Fos/NFATc1 signaling pathway in multiple myeloma. Clinical and Experimental Medicine, 23(5), 1581-1596.

+ Open protocol

+ Expand

Osteoclast Differentiation and Resorption Assay

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

BMCs were seeded on 96-well plates for osteoclast differentiation assay or osteo assay plates for resorption assay at a density of 1.0 × 10⁵ cells per well and cultured in alpha-modified Eagle’s minimum essential medium (α-MEM) with 15% FBS containing 20 ng/mL M-CSF (#315–02; Peprotech, Rocky Hill, NJ, USA) and 50 ng/mL murine soluble recombinant receptor activation of nuclear factor kappa-B ligand (sRANKL, #315–11; Peprotech). After 2 days, BMCs were cultured in the presence of 20 ng/mL M-CSF and 50 ng/mL sRANKL with 10% FBS and 5% mouse serum for 3 days. Sera were prepared from each group of mice. In regard to the neutralization of C5a, 0.5 μg/mL anti-mouse C5a antibody (#MAB21501; R&D) or rat IgG2 isotype control (#MAB006; R&D) was added to the culture medium. After 3 days, differentiated osteoclasts were identified by TRAP staining and TRAP-positive multinucleated (more than three nuclei) cell numbers were evaluated. Resorption areas were measured by ImageJ after removal of the cells with 1 M NH₄OH.

Munenaga S., Ouhara K., Hamamoto Y., Kajiya M., Takeda K., Yamasaki S., Kawai T., Mizuno N., Fujita T., Sugiyama E, & Kurihara H. (2018). The involvement of C5a in the progression of experimental arthritis with Porphyromonas gingivalis infection in SKG mice. Arthritis Research & Therapy, 20, 247.

+ Open protocol

+ Expand

Molecular Mechanisms of Osteoclastogenesis

Cited 1 time

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

+ Open protocol

+ Expand

Quantification of sRANKL and OPG

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

sRANKL and OPG were quantified in cell conditioned media using the R&D System ELISA kits, MTR00 and MOP00 respectively, according to the manufacturer's instructions. Titration of free sRANKL was computed by the difference between equivalent weight of sRANKL and OPG obtained from ELISA, assuming 1:1 as reactive normality of sRANKL:OPG ratio.
Human recombinant sRANKL (Peprotech, cat # 310-01) was used for the in vitro osteoclastogenic assay.

Cappariello A., Paone R., Maurizi A., Capulli M., Rucci N., Muraca M, & Teti A. (2015). Biotechnological approach for systemic delivery of membrane Receptor Activator of NF-κB Ligand (RANKL) active domain into the circulation. Biomaterials, 46, 58-69.

+ Open protocol

+ Expand

Osteoclast Differentiation from PBMCs

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

PBMCs were isolated by density gradient centrifugation and plated in 96-well culture plates at a density of 7.0x10⁵ cells/well and in 24-well culture plates at a density of 1.5x10⁶ cells/well in Dulbecco's Modified Eagle Medium (DMEM; Invitrogen) supplemented with 5000 U Penicilin/Streptomicin (Invitrogen), 2 mM L-Glutamine (Invitrogen) and 10% Fetal Bovine Serum (FBS; Invitrogen) and incubated in a humidified atmosphere at 37°C, 5% CO₂. PBMCs were left overnight for OC precursors (OCPs) to adhere on bone slices. On the following day (day 1 of culture) medium was changed to DMEM supplemented with M-CSF 25 ng/mL (Peprotech) and three days later, medium was again changed to DMEM supplemented with M-CSF (25 ng/mL), sRANKL (50 ng/mL; Peprotech), dexamethasone (10 nM; Sigma Aldrich) and TGF-β (2.5 ng/mL; R&D Systems) in order to differentiate the osteoclast precursors into mature osteoclasts. The culture medium was then changed twice a week. Adherent cells at day 1 and cells cultured on bone slices for 7, 14 and 21 days [13 (link)] were used for functional assays and gene expression.

Perpétuo I.P., Raposeiro R., Caetano-Lopes J., Vieira-Sousa E., Campanilho-Marques R., Ponte C., Canhão H., Ainola M, & Fonseca J.E. (2015). Effect of Tumor Necrosis Factor Inhibitor Therapy on Osteoclasts Precursors in Ankylosing Spondylitis. PLoS ONE, 10(12), e0144655.

+ Open protocol

+ Expand

Osteoclastogenesis Assay with RAW 264.7

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

Osteoclastogenesis assay using RAW 264.7 cells (ATCC, TIB-71) was performed using filtered (pore size = 0.2 μm) culture supernatant of PBL and human GMC. Briefly, RAW 264.7 cells (10³ cells/well) were cultured with 50% supernatant in a 96-well plate. 100 ng/ml of sRANKL (Peprotech) or 10 ng/ml of TNF-α (Peprotech) were used as positive control. In some experiments, rabbit IgG anti-RANKL antibody, OPG-Fc (a kind gift from Dr. Colin Dunstan, Amgen Inc., Thousand Oaks CA), or goat IgG anti-TNF-α antibody (Peprotech) was applied. After cultivation, RNAs were extracted in some experiments to detect osteoclast differentiation marker gene transcripts by RT-PCR, or cells were stained for tartrate-resistant acid phosphatase (TRAP) using Acid Phosphatase kit (Sigma-Aldrich) according to the manufacturer’s instructions. Dark red multinucleated cells (over 3 nuclei) were counted as osteoclasts.

Kanzaki H., Makihira S., Suzuki M., Ishii T., Movila A., Hirschfeld J., Mawardi H., Lin X., Han X., Taubman M, & Kawai T. (2016). Soluble RANKL cleaved from activated lymphocytes by TNF-α converting enzyme (TACE) contributes to osteoclastogenesis in periodontitis. Journal of immunology (Baltimore, Md. : 1950), 197(10), 3871-3883.

+ Open protocol

+ Expand

Osteoclast Induction in Raw264.7 Cells

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

Osteoclast induction of Raw264.7 cells was performed as described (Sun et al., 2016), with modifications. Briefly, Raw264.7 cells were seeded into 96‐well plates at a density of 1 × 10⁴ cells/well, allowed to attach for 24 h, and then cultured in the presence or absence of recombinant murine sRANKL (100 ng/ml, PeproTech, USA) in combination with exosomes or miRNAs for 6 days. The medium was changed every 2 days, and after 6 days of culture, osteoclast differentiation was assessed by means of Trap staining and Trap‐activity detection (Beyotime).

Yu L., Sui B., Fan W., Lei L., Zhou L., Yang L., Diao Y., Zhang Y., Li Z., Liu J, & Hao X. (2021). Exosomes derived from osteogenic tumor activate osteoclast differentiation and concurrently inhibit osteogenesis by transferring COL1A1‐targeting miRNA‐92a‐1‐5p. Journal of Extracellular Vesicles, 10(3), e12056.

+ 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!