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    runx2-antibody-epr22858-106-chip-grade-ab236639.pdf

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Epigenetics and Nuclear Signaling Transcription Other factors
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RecombinantRabMAb

Recombinant Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade (ab236639)

  • Datasheet
  • SDS
  • Certificate of Compliance
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ChIC/CUT&RUN sequencing - Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade (ab236639)
  • Western blot - Anti-RUNX2 antibody [EPR22858-106] (ab236639)
  • Western blot - Anti-RUNX2 antibody [EPR22858-106] (ab236639)
  • Immunoprecipitation - Anti-RUNX2 antibody [EPR22858-106] (ab236639)
  • Immunoprecipitation - Anti-RUNX2 antibody [EPR22858-106] (ab236639)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX2 antibody [EPR22858-106] (ab236639)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX2 antibody [EPR22858-106] (ab236639)
  • ChIP - Anti-RUNX2 antibody [EPR22858-106] (ab236639)
  • 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

Conjugates logo Related conjugates and formulations

Carrier Free

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Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Primary
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Anti-RUNX2 antibody [EPR22858-106] - BSA and Azide free (ab264077)
Protein
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ChIC/CUT&RUN pAG-MNase (ab285373)

View more associated products

Overview

  • 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.

  • 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.

  • Positive control

    • WB: PC-3, MDA-MB-231, Saos-2, MEF, C2C12, MCF7, HeLa, C6 and PC-12 lysates. IHC-P: Human osteosarcoma and Rat embryo 14.5 day tissues. IP: PC-3 and Saos-2 cells.
  • 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
    For more information see here.

    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

  • 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...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR22858-106
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Transcription Factors
    • Stem Cells
    • Mesenchymal Stem Cells
    • Osteogenesis
    • Epigenetics and Nuclear Signaling
    • ChIP assays
    • ChIP antibodies
    • Developmental Biology
    • Organogenesis
    • Skeletal development
    • Bone
    • Neuroscience
    • Development

Associated products

  • Alternative Versions

    • Anti-RUNX2 antibody [EPR22858-106] - BSA and Azide free (ab264077)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

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
ChIC/CUT&RUN-seq
Use 5µg for 105 cells.
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.
Notes
ChIC/CUT&RUN-seq
Use 5µg for 105 cells.
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.
Application notes
Is unsuitable for Flow Cyt or ICC/IF.

Target

  • 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
    modifications

    Phosphorylated; probably by MAP kinases (MAPK) (By similarity). Isoform 3 is phosphorylated on Ser-340.
  • Cellular localization

    Nucleus.
  • Target information above from: UniProt accession Q13950 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 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
    • Unigene: 391013 Mouse
    • Unigene: 391017 Mouse
    • Unigene: 214214 Rat
    • Unigene: 83672 Rat
    see all
  • Alternative names

    • Acute myeloid leukemia 3 protein antibody
    • Alpha subunit 1 antibody
    • AML3 antibody
    • CBF alpha 1 antibody
    • CBF-alpha-1 antibody
    • CBFA1 antibody
    • CCD antibody
    • CCD1 antibody
    • Cleidocranial dysplasia 1 antibody
    • Core binding factor antibody
    • Core binding factor runt domain alpha subunit 1 antibody
    • Core binding factor subunit alpha 1 antibody
    • Core-binding factor subunit alpha-1 antibody
    • MGC120022 antibody
    • MGC120023 antibody
    • Oncogene AML 3 antibody
    • Oncogene AML-3 antibody
    • OSF 2 antibody
    • OSF-2 antibody
    • OSF2 antibody
    • Osteoblast specific transcription factor 2 antibody
    • Osteoblast-specific transcription factor 2 antibody
    • OTTHUMP00000016533 antibody
    • PEA2 alpha A antibody
    • PEA2-alpha A antibody
    • PEA2aA antibody
    • PEBP2 alpha A antibody
    • PEBP2-alpha A antibody
    • PEBP2A1 antibody
    • PEBP2A2 antibody
    • PEBP2aA antibody
    • PEBP2aA1 antibody
    • Polyomavirus enhancer binding protein 2 alpha A subunit antibody
    • Polyomavirus enhancer-binding protein 2 alpha A subunit antibody
    • Runt domain antibody
    • Runt related transcription factor 2 antibody
    • Runt-related transcription factor 2 antibody
    • RUNX2 antibody
    • RUNX2_HUMAN antibody
    • SL3 3 enhancer factor 1 alpha A subunit antibody
    • SL3-3 enhancer factor 1 alpha A subunit antibody
    • SL3/AKV core binding factor alpha A subunit antibody
    • SL3/AKV core-binding factor alpha A subunit antibody
    see all

Images

  • ChIC/CUT&RUN sequencing - Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade (ab236639)
    ChIC/CUT&RUN sequencing - Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade (ab236639)

    CUT&RUN was performed using the ChIC/CUT&RUN pAG-MNAse ab285373, 105 HeLa cells and 5µg of Ab236639 [EPR22858-106]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads.

    Additional screenshots of mapped reads can be downloaded here.

  • Western blot - Anti-RUNX2 antibody [EPR22858-106] (ab236639)
    Western blot - Anti-RUNX2 antibody [EPR22858-106] (ab236639)
    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

     

  • Western blot - Anti-RUNX2 antibody [EPR22858-106] (ab236639)
    Western blot - Anti-RUNX2 antibody [EPR22858-106] (ab236639)
    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

  • Immunoprecipitation - Anti-RUNX2 antibody [EPR22858-106] (ab236639)
    Immunoprecipitation - Anti-RUNX2 antibody [EPR22858-106] (ab236639)

    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

     

  • Immunoprecipitation - Anti-RUNX2 antibody [EPR22858-106] (ab236639)
    Immunoprecipitation - Anti-RUNX2 antibody [EPR22858-106] (ab236639)

    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

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX2 antibody [EPR22858-106] (ab236639)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX2 antibody [EPR22858-106] (ab236639)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX2 antibody [EPR22858-106] (ab236639)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RUNX2 antibody [EPR22858-106] (ab236639)

    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.

  • ChIP - Anti-RUNX2 antibody [EPR22858-106] (ab236639)
    ChIP - Anti-RUNX2 antibody [EPR22858-106] (ab236639)

    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

  • Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade (ab236639)
    Anti-RUNX2 antibody [EPR22858-106] - ChIP Grade (ab236639)

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (8)

Publishing research using ab236639? Please let us know so that we can cite the reference in this datasheet.

ab236639 has been referenced in 8 publications.

  • Wu K  et al. RUNX2 promotes vascular injury repair by activating miR-23a and inhibiting TGFBR2. Ann Transl Med 9:363 (2021). PubMed: 33842584
  • Mi B  et al. SARS-CoV-2-induced Overexpression of miR-4485 Suppresses Osteogenic Differentiation and Impairs Fracture Healing. Int J Biol Sci 17:1277-1288 (2021). PubMed: 33867845
  • Li S  et al. Metallothionein 3 Promotes Osteoblast Differentiation in C2C12 Cells via Reduction of Oxidative Stress. Int J Mol Sci 22:N/A (2021). PubMed: 33919218
  • Qu G  et al. Comparison of Osteogenic Differentiation Potential of Human Dental-Derived Stem Cells Isolated from Dental Pulp, Periodontal Ligament, Dental Follicle, and Alveolar Bone. Stem Cells Int 2021:6631905 (2021). PubMed: 33927769
  • Li S  et al. Total Flavonoids of Rhizoma Drynariae Promotes Differentiation of Osteoblasts and Growth of Bone Graft in Induced Membrane Partly by Activating Wnt/ß-Catenin Signaling Pathway. Front Pharmacol 12:675470 (2021). PubMed: 34122101
  • Han H  et al. The lncRNA H19/miR-541-3p/Wnt/ß-catenin axis plays a vital role in melatonin-mediated osteogenic differentiation of bone marrow mesenchymal stem cells. Aging (Albany NY) 13:18257-18273 (2021). PubMed: 34311444
  • Yan L  et al. The umbilical cord mesenchymal stem cell-derived exosomal lncRNA H19 improves osteochondral activity through miR-29b-3p/FoxO3 axis. Clin Transl Med 11:e255 (2021). PubMed: 33463060
  • Wangzhou K  et al. MiR-143-3p Inhibits Osteogenic Differentiation of Human Periodontal Ligament Cells by Targeting KLF5 and Inactivating the Wnt/ß-Catenin Pathway. Front Physiol 11:606967 (2020). PubMed: 33603676

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