Recombinant Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade (ab236639)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22858-106] to RUNX2 - ChIP Grade
- Suitable for: ChIC/CUT&RUN-seq, IP, ChIP, IHC-P, WB
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade
See all RUNX2 primary antibodies -
Description
Rabbit monoclonal [EPR22858-106] to RUNX2 - ChIP Grade -
Host species
Rabbit -
Specificity
ab236639 is not recommended for mouse IHC.
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Tested applications
Suitable for: ChIC/CUT&RUN-seq, IP, ChIP, IHC-P, WBmore details
Unsuitable for: Flow Cyt or ICC/IF -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: PC-3, MDA-MB-231, Saos-2, MEF, C2C12, C6 and PC-12 lysates. IHC-P: Human osteosarcoma and Rat embryo 14.5 day tissues. IP: PC-3 and Saos-2 cells. ChIC/CUT&RUN seq: Saos-2 cell
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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: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22858-106 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab236639 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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ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
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IP |
1/30.
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ChIP |
Use at an assay dependent concentration.
Use at 5μg |
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IHC-P |
1/2000.
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WB |
1/1000. Predicted molecular weight: 57 kDa.
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Notes |
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ChIC/CUT&RUN-seq
Use at an assay dependent concentration. |
IP
1/30. |
ChIP
Use at an assay dependent concentration. Use at 5μg |
IHC-P
1/2000. |
WB
1/1000. Predicted molecular weight: 57 kDa. |
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 [EPR22858-106] - ChIP Grade (ab236639) 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.
Predicted band size: 57 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?Western blot: Anti-RUNX2 antibody [EPR22858-106] (ab236639) 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, ab236639 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, 2.5X10^5 of positive cell line Saos-2 or low expression cell line HeLa were used along with 5µg of ab236639 [EPR22858-106]. 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.
Additional screenshots of mapped reads can be downloaded here. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods. -
All lanes : Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade (ab236639) at 1/1000 dilution
Lane 1 : PC-3 (human prostate adenocarcinoma epithelial cell), whole cell lysate
Lane 2 : MDA-MB-231 (human breast adenocarcinoma epithelial cell), whole cell lysate
Lane 3 : Saos-2 (human osteosarcoma epithelial), whole cell lysate
Lane 4 : MEF (mouse embryonic fibroblast (immortalized)), whole cell lysate
Lane 5 : C2C12 (mouse myoblasts myoblast), whole cell lysate
Lane 6 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate
Lane 7 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 57 kDa
Observed band size: 60,64 kDa why is the actual band size different from the predicted?Lysates were made freshly and used in WB test immediately to minimize protein degradation.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:19419310)
Negative control: MCF-7 and HeLa (PMID: 20591170)
Exposure time: 70 seconds
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All lanes : Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade (ab236639) at 1/1000 dilution
Lane 1 : C6 (rat glial tumor glial cell), whole cell lysate
Lane 2 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 57 kDa
Observed band size: 60,64 kDa why is the actual band size different from the predicted?The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 19419310, 20591170)
Exposure time: 3 mintues
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RUNX2 was immunoprecipitated from 0.35 mg Saos-2 (human osteosarcoma epithelial) whole cell lysate with ab236639 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab236639 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: Saos-2 (human osteosarcoma epithelial) whole cell lysate 10ug
Lane 2: ab236639 IP in Saos-2 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab236639 in Saos-2 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds
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RUNX2 was immunoprecipitated from 0.35 mg PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate with ab236639 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab236639 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: PC-3 whole cell lysate 10ug
Lane 2: ab236639 IP in PC-3 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab236639 in PC-3 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds
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Immunohistochemical analysis of paraffin-embedded human osteosarcoma tissue labeling RUNX2 with ab236639 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human osteosarcoma (PMID: 21731849) is observed. The section was incubated with ab236639 for 15 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
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Immunohistochemical analysis of paraffin-embedded rat embryo 14.5 day tissue labeling RUNX2 with ab236639 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on the cartilage cells in the rat embryo 14.5 day tissue. The section was incubated with ab236639 for 15 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
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Legend Chromatin was prepared from Saos-2 cells according to the Abcam Dual-X-ChIP protocol. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab236639 (red), or 5 µg of rabbit normal IgG ab172730 (gray) and 20 µl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).
Primers and probes are commercial primers from PMCID: PMC3281617
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (40)
ab236639 has been referenced in 40 publications.
- Lin H et al. Low-intensity pulsed ultrasound enhances immunomodulation and facilitates osteogenesis of human periodontal ligament stem cells by inhibiting the NF-κB pathway. Cell Tissue Bank 24:45-58 (2023). PubMed: 35644018
- 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
- Li C et al. Continuously released Zn2+ in 3D-printed PLGA/β-TCP/Zn scaffolds for bone defect repair by improving osteoinductive and anti-inflammatory properties. Bioact Mater 24:361-375 (2023). PubMed: 36632506
- Ye W et al. miR-30a inhibits the osteogenic differentiation of the tibia-derived MSCs in congenital pseudarthrosis via targeting HOXD8. Regen Ther 21:477-485 (2022). PubMed: 36313394
- Xie S et al. Diminished arachidonate 5-lipoxygenase perturbs phase separation and transcriptional response of Runx2 to reverse pathological ventricular remodeling. EBioMedicine 86:104359 (2022). PubMed: 36395739