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

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.

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

Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR22858-106
Isotype

IgG
Research areas

Alternative Versions
- Anti-RUNX2 antibody [EPR22858-106] - BSA and Azide free (ab264077)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

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.

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

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?

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)

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)

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)

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)

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)

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)

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

Datasheet

ab236639 has not yet been referenced specifically in any publications.

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

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

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Applications

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Domain

Post-translational
modifications

Cellular localization

Database links

Alternative names

Images

Datasheets and documents

References

Customer reviews and Q&As