Recombinant
RabMAb

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

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

    ab264077 is not recommended for mouse IHC.

  • Tested applications

    Suitable for: IP, ChIP, IHC-P, WBmore details
    Unsuitable for: Flow Cyt or ICC/IF
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human RUNX2 aa 250-350. The exact sequence is proprietary.
    Database link: Q13950

  • 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

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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
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.

False

 

 

  • 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.
    • Information by UniProt
    • Database links

    • 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

    • 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?



      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

       

    • 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

    • 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

       

    • 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

       

    • 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.

    • 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.

    • 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

    References

    ab236639 has not yet been referenced specifically in any publications.

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