Overview

  • Product name

  • Description

    Mouse monoclonal to RUNX2
  • Host species

    Mouse
  • Tested applications

    Suitable for: ELISA, ICC/IF, IHC-P, Flow Cyt, IHC-Fr, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Horse, Dog, Pig, Chimpanzee, Rhesus monkey
  • Immunogen

    Recombinant fragment: NPRPSLNSAP SPFNPQGQSQ ITDPRQAQSS PPWSYDQSYP SYLSQMTSPS IHSTTPLSST RGTGLPAITD VPRRISDDDT ATSDFCLWPS TLSKKSQAGA, corresponding to amino acids 251-351 of Human RUNX2 (NP_004339) with tag.

  • Positive control

    • PC12 cell lysate;
  • General notes

    This product was changed from ascites to tissue culture supernatant on 15 May 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab76956 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
ELISA Use at an assay dependent concentration. Detection limit for recombinant tagged RUNX2 is approximately 0.1ng/ml when used as a capture antibody.
ICC/IF Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. PubMed: 21159735
Flow Cyt Use at an assay dependent concentration.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

IHC-Fr Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 57 kDa.

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

  • Anti-RUNX2 antibody (ab76956) at 1 µg/ml + Recombinant tagged human RUNX2 fragment at 0.2 µg

    Secondary
    Goat anti-Mouse IgG (H&L)-HRP at 1/5000 dilution

    Predicted band size: 57 kDa
    Observed band size: 37 kDa
    why is the actual band size different from the predicted?



    Western blot against tagged recombinant protein immunogen. Predicted band size of immunogen is 37 kDa.

    This image was generated using the ascites version of the product.

  • ab76956 at 10µg/ml staining RUNX2 in HeLa cells by Immunofluorescence.

    This image was generated using the ascites version of the product.

  • Overlay histogram showing Saos 2 cells stained with ab76956 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76956, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    This image was generated using the ascites version of the product.

  • ab76956 staining RUNX2 in Mouse dveloping endochondral bone tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with formaldehyde, permeabilized with 0.1% Triton and blocked with 10% Roche Western Blocking for 1 hour at room temperature. Samples were incubated with primary antibody (1/200 in blocking solution) for 16 hours. An undiluted Alexa Fluor® 555-conjugated Donkey polyclonal was used as the secondary antibody.

    This image was generated using the ascites version of the product.

    See Abreview

  • Anti-RUNX2 antibody (ab76956) at 1 µg/ml + PC12 cell lysate at 25 µg

    Secondary
    Goat anti-Mouse IgG (H&L)-HRP at 1/2500 dilution

    Predicted band size: 57 kDa
    Observed band size: 56 kDa why is the actual band size different from the predicted?



    This image was generated using the ascites version of the product.

References

This product has been referenced in:

  • Qi D  et al. Knockdown of lncRNA-H19 inhibits cell viability, migration and invasion while promotes apoptosis via microRNA-143/RUNX2 axis in retinoblastoma. Biomed Pharmacother 109:798-805 (2019). Read more (PubMed: 30551533) »
  • Chen X  et al. Long non-coding RNA XIST promotes osteoporosis through inhibiting bone marrow mesenchymal stem cell differentiation. Exp Ther Med 17:803-811 (2019). Read more (PubMed: 30651866) »
See all 107 Publications for this product

Customer reviews and Q&As

1-10 of 15 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (mandible)
Permeabilization
Yes - 0.1% TX-100
Specification
mandible
Blocking step
BSA and serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted May 09 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Chondrocytes)
Permeabilization
Yes - TritonX100
Specification
Chondrocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.1% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jun 28 2018

Application
Immunohistochemistry (Frozen sections)
Sample
Jaculus jaculus Tissue sections (jerboa metatarsals)
Permeabilization
No
Specification
jerboa metatarsals
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative
Paraformaldehyde

Haydee Gutierrez

Verified customer

Submitted Jun 12 2018

Application
Western blot
Sample
Pig Cell lysate - whole cell (Heart)
Gel Running Conditions
Non-reduced Denaturing (8% gel)
Loading amount
30 µg
Treatment
TGF beta for 72hrs
Specification
Heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Jun 22 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (Bone and Aortic Valve Tissue)
Antigen retrieval step
Enzymatic - Buffer/Enzyme Used: Trypsin-CaCl 0.001mg/mL 30 min 37*C
Permeabilization
No
Specification
Bone and Aortic Valve Tissue
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
10% NBF

Abcam user community

Verified customer

Submitted Jan 24 2013

Application
Western blot
Sample
Human Cell lysate - whole cell (U2-OS)
Loading amount
25 µg
Specification
U2-OS
Gel Running Conditions
Reduced Denaturing (15)
Blocking step
Odyssey buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Oct 18 2012

Question
Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxxxx. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

Read More

Answer

Thank you for calling us and for alerting us to the problem you are experiencing with our product. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

As we discussed on the phone, I am attaching our IHC and our Mouse on Mouse staining protocols for your review. I hope that you find these informative. If you have any questions please to not hesitate to contact me.

I sincerely hope that the suggestions made today, including using a higher secondary antibody dilution, adding 1% BSA to your blocking buffer and washing with TBS + 0.1% Tween-20 do improve the performance of this product. Should you continue to experience any problems with this product please let me know.

This product is covered by our Abpromise guarantee. If we cannot remedy this issue and this is a product that you have purchased within the last six months, we will replace or refund it under our Abpromise guarantee, as you are using it according to specifications listed on our datasheet.

I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

Use our products? Submit an Abreview. Earn rewards!

Read More

Answer

Thank you for your answer.

The order mentioned is over one unit of ab76956 - an anti RUNX2 antibody which is not conjugated. Can you confirm please that we are referring to the same order and antibody?

Secondly, can you confirm that you have already used this antibody successfully in your laboratory? Which antigen retrieval method are you using and what sections are you staining (tissue, species)?

Please understand that the quality of our antibodies is very important to us and we would therefore like to collect the relevant information when a problem appeared.

We are happy to send you a free replacement vial and thank you for your cooperation in resolving this issue.

Read More

Answer

Thank you for your email.

We are very sorry to hear that there was a problem with this antibody. We have been investigating the best way to mark the boxes. I will liaise with my colleagues in this regardsandforwardyour concerns. Thank you for your honest feedback.

I will be pleased to send you a free of charge replacement vial. You mentioned that you have tested the antibody - can you please let us know how exactly you have tested it (what application, samples, protocol)? Indeed, antibodies are very stable proteins and even a week at room temperature does not impair their activity normally. If you can send us the details of the experiment in which this antibody failed, it would be helpful for our internal investigation.

Thank you for your cooperation. We look forward to hear back from you.

Read More

1-10 of 15 Abreviews or Q&A

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