Runx2

Manufactured by OriGene

Sourced in United States

Runx2 is a protein-coding gene that plays a critical role in the regulation of osteoblast differentiation and bone formation. It is a member of the Runt-related transcription factor family and acts as a master regulator of skeletal development.

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Lab products found in correlation

Vector s 1000, by Vector Laboratories (2 mentions) Pk 4000, by Vector Laboratories (1 mentions) S 1000, by Vector Laboratories (1 mentions) Sk 4100, by Vector Laboratories (1 mentions)

5 protocols using runx2

RUNX2 Overexpression in del-RUNT Cells

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RUNX2 transfection was performed using Lenti ORF particles, RUNX2 (Origene Technologies, Rockville, MD, USA), according to the manufacturer’s instruction. Briefly, when del-RUNT cells were at 60% confluence, they were incubated with and without lentiviral particles and polybrene (1000× stock solution (8 mg/mL). After 4 h, the medium was replaced, and protein detection was performed after 3 days. For each condition, three independent experiments were performed.

Deiana M., Dalle Carbonare L., Serena M., Cheri S., Parolini F., Gandini A., Marchetto G., Innamorati G., Manfredi M., Marengo E., Brandi J., Cecconi D., Mori A., Mina M.M., Antoniazzi F., Mottes M., Tiso N., Malerba G., Zipeto D, & Valenti M.T. (2018). New Insights into the Runt Domain of RUNX2 in Melanoma Cell Proliferation and Migration. Cells, 7(11), 220.

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Branched PEI-HA Synthesis and Chondrogenic Plasmid Complexation

Cited 1 time

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Branched PEI-HA was synthesized as previously described,^{6 (link), 9 (link)} using a reductive amination reaction to directly conjugate the hyaluronic acid fragments (6.4kDa) (LifeCore Biomedical, Chaska, MN) to the primary amines of the bPEI (Sigma-Aldrich, St. Louis, MO). The structure was verified with ¹H NMR to ensure correct conjugation as has been described previously.^{6 (link), 9 (link)}Plasmid DNA encoding for RUNX2, SOX5, SOX6, and SOX9 (Origene, Rockville, MD) was expanded using DNA expansion kits according to the manufacturer’s instructions (Qiagen, Venlo, Netherlands), collected, and used directly. For loading into hydrogels, bPEI-HA and DNA were combined drop wise in a constant 7.5:1 Nitrogen:Phosphate (N:P) ratio and allowed to complex in ultrapure (type 1) water (Super-Q Water Purification System, EMD Millipore, Billerica, MA) at room temperature for 30 min before use. After complexation, complexes were lyophilized for 48 hrs in preparation for use in hydrogel loading.

Needham C.J., Shah S.R., Dahlin R.L., Kinard L.A., Lam J., Watson B.M., Lu S., Kasper F.K, & Mikos A.G. (2014). Osteochondral Tissue Regeneration Through Polymeric Delivery of DNA Encoding for the SOX Trio and RUNX2. Acta biomaterialia, 10(10), 4103-4112.

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Immunohistochemical Analysis of Dental Markers

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Tissue sections were deparaffinized following standard procedures. Endogenous peroxidase activity was quenched by 3% hydrogen peroxide for 5 min, and then washed in PBS. Slides were blocked with 5% goat serum (Vector S-1000) for 1 hour at room temperature. The appropriate primary antibody was added and incubated overnight at 4°C, then washed in PBS. Samples were incubated with appropriate biotinylated secondary antibodies (Vector BA-x) for 30 minutes, and washed in PBS. An advidin/biotinylated enzyme complex (Kit ABC Peroxidase Standard Vectastain PK-4000) was added and incubated for 30 minutes and a DAB substrate kit (Kit Vector Peroxidase subtrate DAB SK-4100) was used to develop the color reaction. Antibodies used include Ki67 (Thermo Scientific, dilution 1: 100), Runx2 (Origene, dilution 1: 200), Osteopontin (NIH LF 175, dilution 1: 4000), dentin sialoprotein (DSP, Millipore, dilution 1: 2000), dentin phosphoprotein (DPP, which was generated by the Department of Dental Science for Health Promotion, Division of Cervico Gnathostomatology, Hiroshima University, dilution 1: 2000). Counterstain was performed with hematoxylin after development with a DAB substrate in order to count cells.

Lim W.H., Liu B., Cheng D., Hunter D.J., Zhong Z., Ramos D.M., Williams B.O., Sharpe P.T., Bardet C., Mah S.J, & Helms J.A. (2014). Wnt signaling regulates pulp volume and dentin thickness. Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research, 29(4), 892-901.

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Immunohistochemical Analysis of Osteogenic Factors

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Alkaline phosphatase staining and DAPI staining were performed to evaluate osteogenic factor and to count cells, respectively (14 (link), 15 (link)). For immunostaining, tissue sections were deparaffinized, endogenous peroxidase activity was quenched by 3% hydrogen peroxide then washed in PBS. Slides were blocked with 5% goat serum (Vector S-1000) for 1 hour at room temperature. The appropriate primary antibody was added and incubated overnight at 4°C, then washed in PBS. Samples were incubated with appropriate biotinylated secondary antibodies (Vector BA-x) for 30 minutes, and washed in PBS. An advidin/biotinylated enzyme complex (Kit ABC Peroxidase Standard Vectastain PK-4000) was added and incubated for 30 minutes and a DAB substrate kit (Kit Vector Peroxidase subtrate DAB SK-4100) was used to develop the color reaction. Antibodies used include Runx2 (Origene, dilution 1: 2000), Osteopontin (NIH LF 175, dilution 1: 4000), Fibromodulin (Santa Cruz Biotech, dilution 1: 1000) and β-catenin (Lab Vision, dilution 1:100).

Lim W.H., Liu B., Cheng D., Williams B.O., Mah S.J, & Helms J.A. (2014). Wnt signaling regulates homeostasis of the periodontal ligament. Journal of periodontal research, 49(6), 751-759.

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Immunohistochemical Analysis of Bone Markers

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After depara nization of tissue sections, endogenous peroxidase activity was dampened using 3% hydrogen peroxide and then washed using phosphate buffered saline (PBS). With 5% goat serum (S-1000, Vector, Burlingame, CA), slides were blocked for 60 minutes at room temperature. The assigned primary antibody was added and incubated for one night at 4°C, then washed using PBS. Incubated with biotinylated secondary antibodies (BA-x, Vector) for 30 minutes, samples were washed using PBS. An advidin/biotinylated enzyme complex (PK-4000, Vector) was combined and incubated for 30 minutes. A 3,3' diaminobenzidine substrate kit (SK-4100, Vector) was used to detect color. Antibodies included Osterix (OriGene, Rockville, MA; 1:2000 dilution), Runx2 (OriGene, Rockville, MA;1:2000 dilution), Osteopontin (LF 175, National Institutes of Health, Bethesda, MD; 1:4000 dilution), RANKL (LabVision AB, Värmdö, Sweden; 1:100 dilution) and TGF-beta (TGF-β) (Abcam plc. Waltham, MA; 1:200 dilution).

Lim J., An J., & Lim W.H. (2022). Effects of Selective Alveolar Decortication on the Periodontal Complex.

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