OPG was measured in conditioned media from cultures of cells treated with EGCG using the murine OPG/TNFRSF11B Duo Set (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instructions. RANKL was measured in conditioned media from cultures of cells treated with EGCG using the murine RANKL (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instructions
Murine rankl
Manufactured by R&D Systems
Sourced in United States
Murine RANKL is a recombinant protein that represents the extracellular domain of the mouse Receptor Activator of NF-κB Ligand. It is a key regulator of osteoclast differentiation and activation.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Leucocyte acid phosphatase kit, by Merck Group (2 mentions)
Nunc lab tek 2 8 well chamber slide system, by Thermo Fisher Scientific (2 mentions)
Rankl, by Thermo Fisher Scientific (2 mentions)
β mercaptoethanol, by Thermo Fisher Scientific (2 mentions)
Adiporon, by Merck Group (2 mentions)
Murine m csf, by Thermo Fisher Scientific (1 mentions)
Ab189491, by Abcam (1 mentions)
Ab137037, by Abcam (1 mentions)
Rna interference sequences, by GenePharma (1 mentions)
Nfatc1, by ABclonal (1 mentions)
Gkt137831, by Apexbio (1 mentions)
6 protocols using murine rankl
1
Quantifying OPG and RANKL in EGCG-treated cells
2
Osteoclastogenesis Induction by Mechanical Stimulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Osteoclastogenesis was induced in BMM cells under the application of mechanical loading. Cells were seeded into 6-well Flexwell plates coated with collagen type 1 (#BF-3001C, Flexcell Int. Corporations, Burlington, NC, USA) in a BMM medium supplemented with 20 ng/mL murine M-CSF (#315-02, PeproTech, Hamburg, Germany) and 50 ng/mL murine Rankl (#462-TEC/CF, R&D Systems, Biotechne, Minneapolis, USA) to induce osteoclastogenesis. After seeding, cells were subjected to a mechanical load for 4 h/day on 5 consecutive days. Subsequently, cells were fixed and stained for tartrate-resistant acid phosphatase activity using the Acid Phosphatase, Leukocyte (TRAP) kit from Sigma-Aldrich (#A387, Taufkirchen, Germany). Membranes of the plates were scanned using the TissueFaxs system. Cells containing three or more nuclei were considered as osteoclasts and were counted.
3
Osteoclastogenesis Regulation via Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Ti nanoparticles were obtained from Nanjing Emperor Nano Materials Company. RNA interference sequences were purchased from GenePharma (Suzhou, China). Murine RANKL was purchased from R&D Systems (Minneapolis, USA). GKT137831 was purchased from ApexBio (Boston, USA). Dulbecco’s modified Eagle’s medium (DMEM/high glucose) was purchased from VivaCell (Shanghai, China) and fetal bovine serum (FBS) was obtained from HyClone (Logan, USA). CCK-8 assay kits were obtained from ApexBio (Boston, USA). The primary antibodies used in our study included NFATc1 (A1539, ABclonal, Wuhan, China), MMP-9 (A0289, ABclonal), NOX4 (A11274, ABclonal), HO-1 (ab189491, Abcam Cambridge, UK), SOD2 (ab137037, Abcam), and Nrf2 (A0674, ABclonal). Secondary antibody was purchased from Multisciences (Hangzhou, China).
4
Osteoclastogenesis Modulation by AdipoRon
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For osteoclastogenesis, a total of 1 × 104 RAW 264.7 (ATCC) cells were seeded per well on 24-well plates in 500 μl αMEM containing 5% FCS (Hyclone, GE Healthcare), 1% penicillin-streptomycin (Gibco) as well as 50 μM β-mercaptoethanol (Gibco) and 30 ng/ml murine RANKL (R&D Systems). To study the impact of AdipoRon on osteoclast differentiation, 10 μM AdipoRon (Sigma-Aldrich) were added to the culture medium. Tartrate-resistant acid phosphatase (TRAcP) activity was examined using the leucocyte acid phosphatase kit (Sigma-Aldrich) according to manufacturer’s specifications. TRAcP+ cells with ≥3 nuclei were considered as osteoclasts. In addition, 5 × 103 RAW 264.7 cells were seeded per well in a Nunc Lab-Tek II 8-well Chamber Slide system (ThermoFisher Scientific) in a total of 300 μl osteoclast differentiation medium containing 30 ng/ml RANKL. Fully differentiated osteoclasts were treated with 10 μM AdipoRon for the indicated time points. FACS analysis as well as sorting of mature OCLs for subsequent Western-blot analyses was performed as described previously (Madel et al., 2018 ).
5
Osteoclast Differentiation and Resorption Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mononuclear hematopoietic cell population from the long bone marrow of 12 week old female mice were harvested and cultured as described above at 37 °C, 7% CO2 with the medium being replaced every 2–3 days. For modified conditions, the growth medium was phenol red free α MEM containing 100 Units/mL Penicillin and 100 μg/mL Streptomycin, 10% Charcoal stripped FBS, 150ng/ml M-CSF, and 30ng/ml murine RANKL (R&D System). Dentine disks were cultured for 17 days, TRAP stained44 (link) and the number of resorbing osteoclasts and the amount of resorption per dentine disk were quantified as above.
6
Osteoclastogenesis Protocol with AdipoRon
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For osteoclastogenesis, a total of 1x10 4 RAW 264.7 (ATCC) cells were seeded per well on 24well plates in 500 µl αMEM containing 5% FCS (Hyclone, GE Healthcare), 1% penicillinstreptomycin (Gibco) as well as 50 μM β-mercaptoethanol (Gibco) and 30 ng/ml murine RANKL (R&D Systems). To study the impact of AdipoRon on osteoclast differentiation, 10 µM AdipoRon (Sigma-Aldrich) were added to the culture medium. Tartrate-resistant acid phosphatase (TRAcP) activity was examined using the leucocyte acid phosphatase kit (Sigma-Aldrich) according to manufacturer's specifications. TRAcP + cells with ≥3 nuclei were considered as osteoclasts. In addition, 5x10 3 RAW 264.7 cells were seeded per well in a Nunc
Lab-Tek II 8-well Chamber Slide system (ThermoFisher Scientific) in a total of 300 µl osteoclast differentiation medium containing 30 ng/ml RANKL. Fully differentiated osteoclasts were treated with 10 µM AdipoRon for the indicated time points. FACS analysis as well as sorting of mature OCLs for subsequent Western-blot analyses was performed as described previously (Madel et al., 2018) (link).
Lab-Tek II 8-well Chamber Slide system (ThermoFisher Scientific) in a total of 300 µl osteoclast differentiation medium containing 30 ng/ml RANKL. Fully differentiated osteoclasts were treated with 10 µM AdipoRon for the indicated time points. FACS analysis as well as sorting of mature OCLs for subsequent Western-blot analyses was performed as described previously (Madel et al., 2018) (link).
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!