Opg elisa kits

Recombinant human BMP2, M-CSF, and RANKL were purchased from PeproTech (Rocky Hill, NJ, USA). Phospho-specific (Ser151) and pan-CRTC1 antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). HG-9-91-01 was purchased from MedChemExpress (Princeton, NJ, USA). Antibodies against SIK1, Id1, Lamin B, and α-tubulin, were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). OCN antibody was acquired from Abcam (Cambridge, UK). Anti-β-actin and anti-HA were acquired from Sigma-Aldrich (St. Louis, MO, USA). HRP Anti-Rabbit IgG (Peroxidase) Polymer Detection Kit was purchase from Vector Laboratories (Burlingame, CA, USA). Additionally, an alkaline phosphatase (ALP) assay kit was purchased from Takara Bio Inc. (Ohtsu, Japan), while an ALP staining kit, tartrate-resistant acid phosphatase (TRAP) staining kit, β-glycerophosphate, and ascorbic acid were purchased from Sigma-Aldrich (St. Louis, MO, USA). Mouse RANKL and OPG ELISA kits were purchased from R&D Systems (Minneapolis, MN, USA). pcDNA3-SIK1-HA, pcDNA3-SIK1-FLAG, and pcDNA3-SIK1-T182A-FLAG plasmids were as previously described^{39 (link)}. pGL-Id1 (−1231/+88) plasmid was provided by Dr. Korchynskyi (National Academy of Sciences in Ukraine)^{40 (link)}. siRNA oligonucleotides for control, SIK1, SIK2, SIK3, or CRTC1 were purchased from Thermo Fisher Scientific (Waltham, MA, USA).

MC3T3-E1 cells with or without LiCl or ICG-001 pretreatment were incubated with 0.1 mg/ml Ti particles at 37 °C for 24 h. The culture medium was harvested for ELISA analysis. Mouse RANKL and OPG ELISA kits (R&D Systems, Shanghai, China) were used to determine the levels of the two cytokines according to the manufacture’s instruction.