Recombinant
RabMAb

Recombinant Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (ab92336)

Overview

  • Product name
    Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099]
    See all RUNX1 / AML1+RUNX3+RUNX2 primary antibodies
  • Description
    Rabbit monoclonal [EPR3099] to RUNX1 / AML1 + RUNX3 + RUNX2
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human RUNX1/ AML1 (C terminal). The exact sequence is proprietary.
    (Peptide available as ab177141)

  • Positive control
    • WB: MOLT4, WEHI-3, CTLL-2 and Raw264.7 cell lysate; mouse and rat thymus tissue lysate, mouse spleen tissue lysate and fetal thymus tissue lysate. IHC: Human tonsil tissue. IP: Molt-4 cell lysate
  • 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 ab92336 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
WB 1/5000 - 1/10000. Predicted molecular weight: 49 kDa.Can be blocked with RUNX1 / AML1 peptide (ab177141).
IP 1/50.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

The use of an HRP/AP polymerized secondary antibody will give a stronger signal.

Flow Cyt 1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/100 - 1/250.

Target

Images

  • All lanes : Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (ab92336) at 1.28 µg/ml (purified)

    Lane 1 : Raw264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
    Lane 2 : Molt-4 (Human lymphoblastic leukemia T lymphoblast) whole cell lysate
    Lane 3 : WEHI-3 (Mouse leukemia lymphoblast) whole cell lysate
    Lane 4 : Mouse thymus lysate
    Lane 5 : CTLL-2 (Mouse T lymphocyte) whole cell lysate
    Lane 6 : Rat thymus lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 0.05 µg/ml

    Predicted band size: 49 kDa



    Blocking/Diluting buffer and concentration: 5% NFDM /TBST 

  • ab92336 (purified) at 1/500 immunoprecipitating RUNX1 / AML1 + RUNX3 + RUNX2 in 10 μg Molt-4 (Human lymphoblastic leukemia T lymphoblast)whole cell lysate (Lanes 1 and 2, observed at 49 kDa). Lane 3 - Rabbit monoclonal IgG (ab172730) instead of ab92336 in Molt-4 whole cell lysate. For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

    Blocking/Dilution buffer and concentration: 5% NFDM/TBST.

  • All lanes : Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (ab92336) at 1.28 µg/ml (purified)

    Lane 1 : Mouse spleen lysate
    Lane 2 : Mouse thymus lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 0.05 µg

    Predicted band size: 49 kDa



    Blocking/Diluting buffer and concentration: 5% NFDM /TBST

  • RUNX2 recombinant protein full length, with N-terminal HIS tag, expressed in E.Coli.

    RUNX3 overexpression and empty vector control lysates created in HEK293T cells.  The protein contains a C-terminal DDK tag.

  • All lanes : Anti-RUNX1 / AML1 + RUNX3 + RUNX2 antibody [EPR3099] (ab92336) at 1/10000 dilution

    Lane 1 : MOLT4
    cell lysate
    Lane 2 : fetal thymus lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit HRP at 1/2000 dilution

    Predicted band size: 49 kDa

  • Immunohistochemistry staining of RUNX1 / AML1 in formalin-fixed, paraffin-embedded Human tonsil tissue using 1/100 ab92336.
  • Flow cytometric analysis of permeabilized Molt-4 cells using anti-RUNX1 ab92336 (red) or a rabbit IgG (negative) (green).
  • ab92336 staining RUNX1 / AML1 in human glioblastoma cell line by Immunocytochemistry/ Immunofluorescence.
    Cells were fixed in paraformaldehyde, permeabilized using 0,1% Triton X 100 in PBS, blocked with 0.5% BSA for 30 minutes at room temperature and then incubated with ab92336 at a 1/50 dilution for 16 hours at 4°C. The secondary used was an Alexa-Fluor 488 conjugated goat anti-rabbit polyclonal used at a 1/400 dilution. Nuclei are counterstained with DAPI.

    See Abreview

  • ab92336 staining RUNX1 / AML1 in rat glioblastoma cell line C6 by Immunocytochemistry/ Immunofluorescence.
    Cells were fixed in paraformaldehyde, permeabilized using 0,1% Triton X 100 in PBS, blocked with 0.5% BSA for 30 minutes at room temperature and then incubated with ab92336 at a 1/50 dilution for 16 hours at 4°C. The secondary used was a Cy3 conjugated goat anti-rabbit polyclonal used at a 1/400 dilution. Nuclei are counterstained with DAPI.

    See Abreview

References

This product has been referenced in:
  • Draper JE  et al. A novel prospective isolation of murine fetal liver progenitors to study in utero hematopoietic defects. PLoS Genet 14:e1007127 (2018). WB ; Mouse . Read more (PubMed: 29300724) »
  • Bergiers I  et al. Single-cell transcriptomics reveals a new dynamical function of transcription factors during embryonic hematopoiesis. Elife 7:N/A (2018). Read more (PubMed: 29555020) »
See all 27 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Question
Answer

Merci de nous avoir contactés.

J'ai donc comme convenu vérifié les séquences immunogènes et les alignements de séquences (voir fichiers joints)pour les deux monoclonaux de lapin anti-RUNX1 / AML1:

ab92336: 78% d'homologie avec la protéine RUNX1 de poulet (http://www.ncbi.nlm.nih.gov/protein/669127)
ab133261: 57% d'homologie avec la protéine RUNX1 de poulet (http://www.ncbi.nlm.nih.gov/protein/669127)



L'anticorps ab92336 (https://www.abcam.com/ab92336) me semble donc êtrele plus approprié pour reconnaitre la protéine de poulet.
Les séquences immunogènes sont des informations confidentielles que nous vous demandons de ne pas divulguer lors de publication ou présentations orales de vos travaux de recherche. Merci de votre compréhension.

Je pense que vous souhaitez participer à notre offre de test etutiliser ab92336 chez le poulet. Le code de réduction correspondant est ****. La date d'expiration de ce code est le jj mm aaaa. Ce code est valablepour l’achat d’un autre anticorps primaire. Inutile de mentionner ce code sur votre première commande de ab92336.

Pour activer ce code, vous devez dans un premier temps soumettre une Abreview avec vos résultatschez le pouletet noter ce code dans la section « Additional notes ». Ainsi activé, ce code vous permettra par la suite de passer votre prochaine commande. Veuillez contacter notre service clientèle en vous munissant des informations suivantes : code de réduction, numéro de commande de ab92336.

Nous restons à votre disposition pour plus d’informations concernant cette offre promotionnelle. Dans l’attente de recevoir votre Abreview nous vous souhaitons bonne chance dans vos recherches.

Termes et Conditions : https://www.abcam.com/collaborationdiscount.

J'espère que ces informations vous seront utiles. N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.

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Question
Answer

Merci pour votre appel.

J'ai fait l'alignement de l-ab92336 avec la séquence de poulet, mais c'est pas super. Veuillez voir vous-même: http://www.ebi.ac.uk/Tools/services/web_clustalo/toolresult.ebi?tool=clustalo&jobId=clustalo-I20120913-111634-0397-65418468-oy

J'ai fait donc aussi l'alignement pour l'autre anticorps monoclonal de lapin contre le RUNX1 que nous avons dans le catalogue, le ab133261. Cet anticorps est testé en Western blot et sur coupe paraffine. L'alignement de l'immunogène avec la séquence du poulet est mieux conservé, veuillez voir le résultat ici: http://www.ebi.ac.uk/Tools/services/web_clustalo/toolresult.ebi?tool=clustalo&jobId=clustalo-I20120913-112516-0204-25979061-pg

Je peux donc vous conseiller le ab133261. https://www.abcam.com/index.html?datasheet=133261 (or use the following: https://www.abcam.com/index.html?datasheet=133261).

Veuillez me confirmer si vous préférez aussi cet anticorps ou si vous souhaitez que nous cherchons pour une autre alternative.

Voici aussi le lien pour les détails pour l'offre de test: https://www.abcam.com/test-vial

Je me réjouis de votre réponse. N'hésitez pas à me contacter aussi pour des questions.

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Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been succesful.

I would like to reassure you that this antibody is tested and covered by our guarantee for WB, IP, IHC-P, ICC, Flow Cyt, ICC/IF and Mouse, Rat, Human samples. Before deciding how to proceed, I would like to investigate this particular case further for you, and also obtain some further information for our quality records.

In order to do this, I have enclosed some questionnaires below. I would appreciate if you could select the correct questionare for the appilcation you are using and complete this. It will help you put the information we require together very easily.

In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund. Alternatively, we can provide an alternative replacement.

Thank you for your time and cooperation. We look forward to receiving the completed questionaire.

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human glioblastoma cell line U87MG)
Specification
human glioblastoma cell line U87MG
Fixative
Paraformaldehyde
Permeabilization
Yes - 1% Triton X 100 in PBS
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 0.5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Feb 06 2012

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (glioblastoma cell line from rat)
Specification
glioblastoma cell line from rat
Fixative
Paraformaldehyde
Permeabilization
Yes - 1% Triton X 100 in PBS
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 0.5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Feb 06 2012

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (glioblastoma cell line C6)
Specification
glioblastoma cell line C6
Fixative
Paraformaldehyde
Permeabilization
Yes - 0,1% Triton X100 in PBS
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 0.5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jan 24 2012

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human glioblastoma cell line)
Specification
human glioblastoma cell line
Fixative
Paraformaldehyde
Permeabilization
Yes - 0,1% Triton X100 in PBS
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 0.5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jan 24 2012

Answer

Thank you for bringing this to our attention.  I have forwarded your data to our lab for their input.  We may end up submitting a recall for this product, but I would like for them to review your results.  I have refunded your purchase. I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways: (1) Redeemed against the original invoice if this hasn't already been paid. (2) Held on the account for use against a future order. (3) A full refund can be offered where no other invoices are outstanding. Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge. To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website. The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB. I hope this experience will not prevent you from purchasing other products from us in the future. I will let you know what the lab has to say.  Our Scientific Support team is always at your service should you require further expert advice    

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