RUNX2/CBFA1 Antibody - BSA Free

Immunohistochemistry-Paraffin: RUNX2/CBFA1 Antibody - BSA Free [NBP1-77461] - RUNX2/CBFA1 Antibody [NBP1-77461] - Vascularization of mineralized cell-laden collagen and interaction with prostate cancer cells. HUVECs formed endothelial networks that were supported by SMA-expressing hMSCs (scale bar: 50m) and were also positive for CD31 (scale bar: 400m). The remainder of hMSCs expressed RUNX2 as a marker for osteogenic differentiation (scale bar: 50m). Rapid fabrication of vascularized and innervated cell-laden bone models with biomimetic intrafibrillar collagen mineralization. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31388010/) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: RUNX2/CBFA1 Antibody [NBP1-77461] - HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with RUNX2/CBFA1 Antibody (NBP1-77461) at 1 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Immunohistochemistry-Paraffin: RUNX2/CBFA1 Antibody - BSA Free [NBP1-77461] - RUNX2/CBFA1 Antibody [NBP1-77461] - Characterization of TLC cultured in chondrogenic pellets. Gene expression of TLC cultured four weeks in chondrogenic pellets or in cell monolayer in control medium: levels of expression of osteoblast markers (1), chondrocyte markers (2) and tenocyte markers (3) (N = 3) assessed by qPCR.* =0.05. Rotator Cuff Tenocytes Differentiate into Hypertrophic Chondrocyte-Like Cells to Produce Calcium Deposits in an Alkaline Phosphatase-Dependent Manner. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31561454/) licensed under a CC-BY license.

Immunohistochemistry: RUNX2/CBFA1 Antibody [NBP1-77461] - Staining of RUNX2 in mouse prostate using DAB with hematoxylin counterstain.

Immunohistochemistry: RUNX2/CBFA1 Antibody - BSA Free [NBP1-77461] - Histological characterization of rotator cuff calcifications (N = 5). (A) Calcific deposits assessed by micro-computed tomography. (B) HE (hematoxylin & eosin) staining of decalcified samples (1 image for each patient). (C) Von Kossa staining on not decalcified samples & characterization of the fibrocartilaginous area with alcian blue staining, SOFG (Safranin O/Fast Green) staining, & immunohistochemical staining for COL2 (brown) with Gill hematoxylin counterstain. (D,E) Representative immunohistochemical staining for ENPP1 (ectonucleotide pyrophosphatase/phosphodiesterase 1), TNAP (tissue non-specific alkaline phosphatase), Runx2, Sox9, Col10 & CD31 (brown). TF: tendon fibers; CD: calcium deposits, FM: fibrocartilaginous metaplasia; OM: osseous metaplasia; CT: connective tissue; V: vessels. Scale bar = 200 µM. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31561454), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu, Mu
• Applications: WB, ICC/IF, IHC
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1 mg/ml

RUNX2/CBFA1 Antibody - BSA Free Summary

• Description: Novus Biologicals Rabbit RUNX2/CBFA1 Antibody - BSA Free (NBP1-77461) is a polyclonal antibody validated for use in IHC, WB and ICC/IF. Anti-RUNX2/CBFA1 Antibody: Cited in 17 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Immunogen: This RUNX2/CBFA1 Antibody was prepared from a synthetic peptide made to an internal region of the human RUNX2 protein (within residues 300-375). [Swiss-Prot Q13950]
• Localization: Nucleus
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: RUNX2
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 1:100
  • Immunohistochemistry 1:200
  • Immunohistochemistry-Paraffin 1:200
  • Western Blot reported in scientific literature (PMID 29208768)
• Application Notes: Prior to immunostaining paraffin tissues antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.
• Theoretical MW: 56.6 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS and 30% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for RUNX2/CBFA1 Antibody - BSA Free

• Acute myeloid leukemia 3 protein
• CBFA1
• CBF-alpha-1
• CCD1
• CCDAML3
• CLCD
• Core-binding factor subunit alpha-1
• core-binding factor, runt domain, alpha subunit 1
• MGC120023
• ML3
• oncogene AML-3
• OSF2
• OSF-2
• osteoblast-specific transcription factor 2
• PEA2aA
• PEA2-alpha A
• PEBP2A
• PEBP2aA
• PEBP2-alpha A
• polyomavirus enhancer-binding protein 2 alpha A subunit
• runt domain, alpha subunit 1
• runt related transcription factor 2
• runt-related transcription factor 2
• RUNX2
• SL3/AKV core-binding factor alpha A subunit
• SL3-3 enhancer factor 1 alpha A subunit

Background

Runt-related transcription factor 2 (RUNX2), also known as CBFA1, AML-3, PEBP-2alphaA, and OSF-2, is a transcription factor that places a critical role in osteoblast differentiation and bone development (1-3). RUNX2 is a DNA-binding protein that belongs to the RUNX family which share a common runt domain (3). RUNX2 has two main isoforms which vary based on the two promoter regions (3). The main canonical isoform (P1) has MASN/DS at its N-terminus while the other (P2) isoform includes a MRIPV pentapeptide at its N-terminus (3). The RUNX2 P1 isoform has a theoretical molecular weight of 56 kDa and is synthesized as a 521 amino acid (aa) protein containing multiple domains. Specifically, RUNX2 contains transactivation domains (AD1, 2 and 3), a glutamine/alanine (Q/A)-rich domain, a runt homology domain (RHD), a nuclear localization signal (NLS), a proline/serine/threonine (PST)-rich domain, a nuclear matrix targeting signal (NMTS), a repression domain (RD), and a VWRPY region (3). RUNX2 is a heterodimer of an alpha and beta subunit where the alpha subunit binds DNA through the runt domain and the binding affinity is increased through heterodimerization (4).

Functionally, RUNX2 promotes the expression of osteoblast-specific genes vital for the osteoblast differentiation and proliferation process including type I collagen, osteocalcin (OCN), and alkaline phosphatase (APC) (1, 3). Further evidence for the role of RUNX2 is highlighted by a study of Runx2-/-mice which completely lack osteoblasts (4). Additionally, RUNX2 is also required for chondrocyte maturation, which are the cells responsible for cartilage formation (1, 3, 5). Given the role of RUNX2 in bone and cartilage maturation and formation, it is clear that defects or mutations in RUNX2 cause various bone and bone-related diseases (3, 6, 7). For instance, cleidocranial dysplasia (CCD), which presents with delayed cranial suture closure phenotypes, hypoplastic clavicles, extra teeth, and short stature, is caused by haploinsufficiency in RUNX2 (2, 3, 6). Furthermore, metaphyseal dysplasia with maxillary hypoplasia and brachydactyly (MDMHB) is a bone dysplasia disorder with a phenotype of abnormalities in the long bones, an underdeveloped jawbone, and short fingers that is caused by a duplication in RUNX2 (6). Finally, RUNX2 has been shown to be upregulated in mouse models of the joint disorder osteoarthritis (OA) and may be a potential molecular target for disease treatment (7).

Alternative names for RUNX2 include Acute myeloid leukemia 3 protein CBFA1, CBF-alpha-1, CCD1, CCDAML3, CLCD, Core-binding factor subunit alpha-1, MGC120023, ML3, oncogene AML-3, OSF2, osteoblast-specific transcription factor 2, PEA2aA, PEA2-alpha A, PEBP2A, polyomavirus enhancer-binding protein 2 alpha A subunit, runt related transcription factor 2, SL3/AKV core-binding factor alpha A subunit, and SL3-3 enhancer factor 1 alpha A subunit.

References

1. Ferreira, L. B., Gimba, E., Vinagre, J., Sobrinho-Simoes, M., & Soares, P. (2020). Molecular Aspects of Thyroid Calcification. International journal of molecular sciences. https://doi.org/10.3390/ijms21207718

2. Kim, W. J., Shin, H. L., Kim, B. S., Kim, H. J., & Ryoo, H. M. (2020). RUNX2-modifying enzymes: therapeutic targets for bone diseases. Experimental & molecular medicine. https://doi.org/10.1038/s12276-020-0471-4

3. Vimalraj, S., Arumugam, B., Miranda, P. J., & Selvamurugan, N. (2015). Runx2: Structure, function, and phosphorylation in osteoblast differentiation. International journal of biological macromolecules. https://doi.org/10.1016/j.ijbiomac.2015.04.008

4. Uniprot (Q13950)

5. Komori T. (2017). Roles of Runx2 in Skeletal Development. Advances in experimental medicine and biology. https://doi.org/10.1007/978-981-10-3233-2_6

6. Moffatt, P., Ben Amor, M., Glorieux, F. H., Roschger, P., Klaushofer, K., Schwartzentruber, J. A., Paterson, A. D., Hu, P., Marshall, C., FORGE Canada Consortium, Fahiminiya, S., Majewski, J., Beaulieu, C. L., Boycott, K. M., & Rauch, F. (2013). Metaphyseal dysplasia with maxillary hypoplasia and brachydactyly is caused by a duplication in RUNX2. American journal of human genetics. https://doi.org/10.1016/j.ajhg.2012.12.001

7. Chen, D., Kim, D. J., Shen, J., Zou, Z., & O'Keefe, R. J. (2019). Runx2 plays a central role in Osteoarthritis development. Journal of orthopaedic translation. https://doi.org/10.1016/j.jot.2019.11.008

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for RUNX2/CBFA1 Antibody (NBP1-77461)(18)

Publications using NBP1-77461

• Shi L, He Z, Hiraga T et al. Identification of novel macrophages and bone morphogenetic protein signals causing ectopic calcification and impairing muscle regeneration iScience 2025-07-18 [PMID: 40612505]
• Sousa MGDC, Balbinot GS, Subbiah R et al. In vitro development and optimization of cell-laden injectable bioprinted gelatin methacryloyl (GelMA) microgels mineralized on the nanoscale bioRxiv : the preprint server for biology 2023-10-12 [PMID: 37873385] (Immunocytochemistry/ Immunofluorescence, Mouse)
• Chan, B;Glogauer, M;Wang, Y;Wrana, J;Chan, K;Beier, F;Bali, S;Hinz, B;Parreno, J;Ashraf, S;Kandel, R; Adseverin, an actin-binding protein, modulates hypertrophic chondrocyte differentiation and osteoarthritis progression Science advances 2023-08-04 [PMID: 37540756] (Immunocytochemistry/ Immunofluorescence, Human)
• Poudel SB, Ruff RR, Yildirim G et al. Excess growth hormone triggers inflammation-associated arthropathy, subchondral bone loss, and arthralgia The American journal of pathology 2023-03-02 [PMID: 36870529] (IHC-P, Mouse)
• Subbiah R, Lin EY, Athirasala A et al. Engineering of an osteoinductive and growth factor-free injectable bone-like microgel for bone regeneration Advanced healthcare materials 2023-02-20 [PMID: 36808718]
• Zanut A, Li R, Deng R et al. A polymer canvas with the stiffness of the bone matrix to study and control mesenchymal stem cell response Advanced healthcare materials 2022-12-24 [PMID: 36565136]
• Ciavarella C, Motta I, Vasuri F et al. Involvement of miR-30a-5p and miR-30d in Endothelial to Mesenchymal Transition and Early Osteogenic Commitment under Inflammatory Stress in HUVEC Biomolecules 2021-02-05 [PMID: 33562690] (ICC/IF, Human)
• Serjeant M, Moon PM, Quinonez D, et al. The Role of Panx3 in Age-Associated and Injury-Induced Intervertebral Disc Degeneration International journal of molecular sciences 2021-01-22 [PMID: 33499145] (IF/IHC, Mouse)
• Cahill SV, Kwon HK, Back J et al. Locally-delivered Adjuvant Biofilm-penetrating Antibiotics Rescue Impaired Endochondral Fracture Healing Caused by MRSA Infection Journal of orthopaedic research : official publication of the Orthopaedic Research Society 2020-12-18 [PMID: 33336805]
• Chen PY, Qin L, Li G et al. Smooth Muscle Cell Reprogramming in Aortic Aneurysms Cell Stem Cell 2020-04-02 [PMID: 32243809] (Mouse)
• Darrieutort-Laffite C, Arnolfo P, Garraud T et al. Rotator Cuff Tenocytes Differentiate into Hypertrophic Chondrocyte-Like Cells to Produce Calcium Deposits in an Alkaline Phosphatase-Dependent Manner J Clin Med [PMID: 31561454] (IF/IHC)
• Thrivikraman G, Athirasala A, Gordon R et al. Rapid fabrication of vascularized and innervated cell-laden bone models with biomimetic intrafibrillar collagen mineralization Nat Commun 2019-08-06 [PMID: 31388010] (ICC/IF, Human)
• Priante G, Ceol M, Gianesello L et al. Human proximal tubular cells can form calcium phosphate deposits in osteogenic culture: role of cell death and osteoblast-like transdifferentiation. Cell Death Discov 2019-01-28 [PMID: 30701089] (WB, Human)
• Priante G, Quaggio F, Gianesello L et al. Caspase-independent programmed cell death triggers Ca2PO4 deposition in an in vitro model of nephrocalcinosis. Biosci. Rep. 2017-12-05 [PMID: 29208768] (WB, Human)
• Pillai IC, Li S, Romay M et al. Cardiac Fibroblasts Adopt Osteogenic Fates and Can Be Targeted to Attenuate Pathological Heart Calcification. Cell Stem Cell. 2016-11-15 [PMID: 27867037]
• Mao AS, Shin JW, Utech S et al. Deterministic encapsulation of single cells in thin tunable microgels for niche modelling and therapeutic delivery. Nat Mater. 2016-10-31 [PMID: 27798621]
  
  Details:
  This citation used the Alexa Fluor 647 version of this antibody.
• Mao AS, Shin JW, Mooney DJ. Effects of substrate stiffness and cell-cell contact on mesenchymal stem cell differentiation. Biomaterials. 2016-05-05 [PMID: 27203745] (ICC/IF, Mouse)
  
  Details:
  This publication used the AF647 conjugated form of this antibody (NBP1-77461AF647)
• Al-Rekabi Z, Wheeler MM, Leonard A et al. Activation of the IGF1 Pathway Mediates Changes in Cellular Contractility and Motility in Single-Suture Craniosynostosis J Cell Sci 2015-12-11 [PMID: 26659664] (ICC/IF, Human)
  
  Details:
  This citation used the DyLight 488 version of this antibody.

FAQs for RUNX2/CBFA1 Antibody (NBP1-77461). (Showing 1 - 2 of 2 FAQs).

1. We would like an anti-RUNX2 for IHC-P which share cross reactivity with Rat, but not with Human.
   • We don't have any data for our RUNX2 antibodies that confirms they will NOT detect the human protein. When we can confirm that an antibody will not react with a certain species, we display a (-) sign on the datasheet. Otherwise, if the species is not listed it means that it has not been tested.
2. I want to chase Runx2/CBFA1 antibody (NBP1-77461). Does it work with frozen section?
   • Our lab has never tested this antibody in IHC-Fr (frozen sections). NBP1-77461 has only be validated in ICC/IF and IHC-P. Therefore we don't know whether this antibody can produce the positive results in your experiments. However our experiences tell us that, as a general rule, the antibodies that can be used in IHC-P, they are likely to produce the positive results in IHC-Fr after some optimizing experiments. If you indeed use NBP1-77461 in IHC-Fr, you will be automatically qualified for our Innovator's Reward that earn a free antibody from us.

Bioinformatics

• Gene Symbol: RUNX2
• Entrez:
  • 860(Human)
  • 12393(Mouse)
• Uniprot:
  • Q13950()