Cellular proteins were extracted, and western blotting was performed as previously reported (Mao et al., 2017 (link)). Briefly, total proteins were isolated from BMSCs with radioimmunoprecipitation assay (RIPA) buffer (Beyotime Biotechnology, Beijing, China) containing protease inhibitors (Abcam, Cambridge, United Kingdom). Membranes were incubated at 4°C overnight with primary antibodies against RUNX2 (1:2,000; Cell Signaling Technology, Danvers, MA, United States), OCN, OPN, and SORBS1 (1:1,000; Proteintech, Wuhan, China). GAPDH (1:3,000; Cell Signaling Technology, Danvers, MA, United States) was used as the internal control. Appropriate secondary antibodies (1:3,000; Cell Signaling Technology, Danvers, MA, United States) were incubated with the blots at 20–25°C for 1 h. After rinsing with Tris-buffered saline (TBS) containing 0.05% (w/v) Tween-20 (TBST), the signals were detected using an enhanced chemiluminescence (ECL) kit (Millipore, Darmstadt, Germany) and a chemiluminescence system (Bio-Rad Laboratories, Hercules, CA, United States) and analyzed with Image Lab v6.0 software.1
Primary antibodies against runx2
Manufactured by Cell Signaling Technology
Sourced in United States
Primary antibodies against RUNX2 are laboratory reagents used for the detection and analysis of the RUNX2 protein in biological samples. RUNX2 is a transcription factor that plays a crucial role in the regulation of osteoblast differentiation and bone formation. These primary antibodies can be used in various techniques, such as Western blotting, immunohistochemistry, and immunocytochemistry, to study the expression and localization of RUNX2 in cells and tissues.
Automatically generated - may contain errors
Lab products found in correlation
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Appropriate secondary antibody, by Cell Signaling Technology (1 mentions)
Protease inhibitor, by Abcam (1 mentions)
Enhanced chemiluminescence ecl kit, by Merck Group (1 mentions)
Chemiluminescence system, by Bio-Rad (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Ripa buffer, by Beyotime (1 mentions)
Polyvinylidene fluoride membrane, by GE Healthcare (1 mentions)
Col1α1, by Santa Cruz Biotechnology (1 mentions)
Enhanced chemiluminescence kit, by GE Healthcare (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
α tubulin, by Cell Signaling Technology (1 mentions)
Osterix, by Santa Cruz Biotechnology (1 mentions)
Ripa lysis and extraction buffer, by Thermo Fisher Scientific (1 mentions)
2 protocols using primary antibodies against runx2
1
Protein Expression Analysis of BMSCs
2
Osteogenic Protein Expression Analysis
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The hPDLSCs (3.0 × 105 cells/well) were seeded in a 6-well plate and incubated for 2 weeks. Cell pellets were washed with PBS and lysed in ice-cold RIPA lysis and extraction buffer (Thermo Fisher Scientific). Protein concentrations of cell lysates were measured by a Pierce BCA Protein Assay Kit (Thermo Fisher Scientific). Equal amounts of cell extracts (20 μg of protein) were resolved by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and blotted onto a polyvinylidene fluoride membrane (GE Healthcare, Buckinghamshire, UK). After blocking with 5% skim milk, the membranes were incubated with primary antibodies against Runx2, α-tubulin (Cell Signaling, Danvers, MA, USA), Col1α1, and osterix (Santa Cruz Biotechnology, Santa Cruz, CA, USA), followed by incubation with horseradish peroxidase-conjugated secondary antibodies (Thermo Fisher Scientific). Blots were finally visualized using an enhanced chemiluminescence kit (GE Healthcare).
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