Recombinant Anti-RUNX2 antibody [EPR14334] (ab192256)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR14334] to RUNX2
- Suitable for: IHC-P, ICC/IF, Flow Cyt (Intra), ChIC/CUT&RUN-seq
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-RUNX2 antibody [EPR14334]
See all RUNX2 primary antibodies -
Description
Rabbit monoclonal [EPR14334] to RUNX2 -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, ICC/IF, Flow Cyt (Intra), ChIC/CUT&RUN-seqmore details -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- Human osteosarcoma, Human tonsil and Mouse spleen tissues; Saos-2 and PC cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 59% PBS, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR14334 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab192256 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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IHC-P | (5) |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/1000.
For unpurified use at 1/500. |
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Flow Cyt (Intra) |
1/50.
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ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
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Notes |
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IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/1000. For unpurified use at 1/500. |
Flow Cyt (Intra)
1/50. |
ChIC/CUT&RUN-seq
Use at an assay dependent concentration. |
Target
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Function
Transcription factor involved in osteoblastic differentiation and skeletal morphogenesis. Essential for the maturation of osteoblasts and both intramembranous and endochondral ossification. CBF binds to the core site, 5'-PYGPYGGT-3', of a number of enhancers and promoters, including murine leukemia virus, polyomavirus enhancer, T-cell receptor enhancers, osteocalcin, osteopontin, bone sialoprotein, alpha 1(I) collagen, LCK, IL-3 and GM-CSF promoters (By similarity). Inhibits MYST4-dependent transcriptional activation. -
Tissue specificity
Specifically expressed in osteoblasts. -
Involvement in disease
Defects in RUNX2 are the cause of cleidocranial dysplasia (CLCD) [MIM:119600]; also known as cleidocranial dysostosis (CCD). CLCD is an autosomal dominant skeletal disorder with high penetrance and variable expressivity. It is due to defective endochondral and intramembranous bone formation. Typical features include hypoplasia/aplasia of clavicles, patent fontanelles, wormian bones (additional cranial plates caused by abnormal ossification of the calvaria), supernumerary teeth, short stature, and other skeletal changes. In some cases defects in RUNX2 are exclusively associated with dental anomalies. -
Sequence similarities
Contains 1 Runt domain. -
Domain
A proline/serine/threonine rich region at the C-terminus is necessary for transcriptional activation of target genes and contains the phosphorylation sites. -
Post-translational
modificationsPhosphorylated; probably by MAP kinases (MAPK) (By similarity). Isoform 3 is phosphorylated on Ser-340. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 860 Human
- Entrez Gene: 12393 Mouse
- Entrez Gene: 367218 Rat
- Omim: 600211 Human
- SwissProt: Q13950 Human
- SwissProt: Q08775 Mouse
- SwissProt: Q9Z2J9 Rat
- Unigene: 535845 Human
see all -
Alternative names
- Acute myeloid leukemia 3 protein antibody
- Alpha subunit 1 antibody
- AML3 antibody
see all
Images
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All lanes : Anti-RUNX2 antibody [EPR14334] (ab192256) at 1/1000 dilution
Lane 1 : Saos-2 cell lysate
Lane 2 : MC3T3-E1 undifferentiated cell lysate
Lane 3 : MC3T3-E1 7-day Osteogenic differentiation cell lysate
Lane 4 : MC3T3-E1 14-day Osteogenic differentiation cell lysate
Lane 5 : MC3T3-E1 28-day Osteogenic differentiation cell lysate
Lane 6 : C2C12 cell lysate
Lane 7 : SH-SY5Y cell lysate
Lane 8 : NIH/3T3 cell lysate
Lane 9 : LNCaP cell lysate
Lysates/proteins at 20 µg per lane.
Observed band size: 60 kDa why is the actual band size different from the predicted?Western blot: Anti-RUNX2 antibody [EPR14334] (ab192256) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab192256 was shown to bind specifically to RUNX2. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
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ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/µL, 2.5 x 105 Saos-2 cells and 5 µg of ab192256 [EPR14334]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
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ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/µL, 2.5 x 105 Saos-2 cells and 5 µg of ab192256 [EPR14334]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
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ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/µL, 2.5 x 105 Saos-2 cells and 5 µg of ab192256 [EPR14334]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
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Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling RUNX2 with ab192256 at 1/1000 dilution. A ready to use HRP Polymer for Rabbit IgG was used as the secondary. Hematoxylin counterstain.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunocytochemistry/Immunofluorescence analysis of Saos-2 (Human osteosarcoma cell line) labeling RUNX2 with purified ab192256 at 1/1000 dilution. Cells were fixed with 4% PFA and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG (Alexa Fluor®488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.
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Immunohistochemical analysis of paraffin-embedded Human osteosarcoma tissue labeling RUNX2 with ab192256 at 1/1000 dilution. A ready to use HRP Polymer for Rabbit IgG was used as the secondary. Hematoxylin counterstain.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ab192256 staining RUNX2 in PC-3 (human prostate adenocarcinoma) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde, permabilised with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/50. A goat anti rabbit IgG (Alexa Fluorr® 488) at a dilution of 1/2000 was used as the secondary antibody.
Isoytype control: Rabbit monoclonal IgG (Black)
Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
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Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling RUNX2 with ab192256 at 1/1000 dilution. A ready to use HRP Polymer for Rabbit IgG was used as the secondary. Hematoxylin counterstain.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% formaldehyde fixed PC3 cells labeling RUNX2 using ab192256 at a 1/500 dilution. A Goat anti rabbit IgG (Alexa Fluor®488) ab150077 was used as the secondary at a 1/200 dilution. Counterstain DAPI. Permeabilized using 0.1% Triton X-100. The two negative controls: 1. Primary ab concentration (anti-RUNX2) is 1:500 dilution, Secondary ab (Goat anti mouse IgG (Alexa Fluor®594)) is 1:500 dilution; 2. Primary ab concentration (anti-RUNX2) is 1:500 dilution, Secondary ab (Goat anti mouse IgG (Alexa Fluor®594)) is 1:500 dilution.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (92)
ab192256 has been referenced in 92 publications.
- Wu C et al. Mapk7 deletion in chondrocytes causes vertebral defects by reducing MEF2C/PTEN/AKT signaling. Genes Dis 11:964-977 (2024). WB, IHC-Fr ; Mouse . PubMed: 37692479
- Sun B et al. Irisin reduces bone fracture by facilitating osteogenesis and antagonizing TGF-β/Smad signaling in a growing mouse model of osteogenesis imperfecta. J Orthop Translat 38:175-189 (2023). PubMed: 36439629
- Gao P et al. circEXOC5 promotes acute lung injury through the PTBP1/Skp2/Runx2 axis to activate autophagy. Life Sci Alliance 6:N/A (2023). PubMed: 36302650
- Verisqa F et al. Digital Light Processing 3D Printing of Gyroid Scaffold with Isosorbide-Based Photopolymer for Bone Tissue Engineering. Biomolecules 12:N/A (2022). PubMed: 36421706
- Tseng HW et al. Interleukin-1 Is Overexpressed in Injured Muscles Following Spinal Cord Injury and Promotes Neurogenic Heterotopic Ossification. J Bone Miner Res 37:531-546 (2022). PubMed: 34841579