Recombinant Anti-RUNX1 / AML1 antibody [EPR23044-100] (ab240639)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23044-100] to RUNX1 / AML1
- Suitable for: ChIC/CUT&RUN-seq, Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-RUNX1 / AML1 antibody [EPR23044-100]
See all RUNX1 / AML1 primary antibodies -
Description
Rabbit monoclonal [EPR23044-100] to RUNX1 / AML1 -
Host species
Rabbit -
Tested applications
Suitable for: ChIC/CUT&RUN-seq, Flow Cyt (Intra), WB, IP, IHC-P, ICC/IFmore details
Unsuitable for: ChIP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: THP-1, Jurkat and MOLT-4 lysates. IHC-P: Human breast carcinoma and Human breast tissues. ICC/IF: Jurkat cells. Flow Cyt (intra): Jurkat cells. IP: Jurkat cells. ChIC/CUT&RUN-Seq: K-562 cells.
Properties
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23044-100 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab240639 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 |
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ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
5µg |
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Flow Cyt (Intra) |
1/500.
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WB | (5) |
1/1000. Predicted molecular weight: 48 kDa.
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IP |
1/30.
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IHC-P | (4) |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF | (3) |
1/100.
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Notes |
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ChIC/CUT&RUN-seq
Use at an assay dependent concentration. 5µg |
Flow Cyt (Intra)
1/500. |
WB
1/1000. Predicted molecular weight: 48 kDa. |
IP
1/30. |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/100. |
Target
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Function
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, LCK, IL-3 and GM-CSF promoters. The alpha subunit binds DNA and appears to have a role in the development of normal hematopoiesis. Isoform AML-1L interferes with the transactivation activity of RUNX1. Acts synergistically with ELF4 to transactivate the IL-3 promoter and with ELF2 to transactivate the mouse BLK promoter. Inhibits MYST4-dependent transcriptional activation. -
Tissue specificity
Expressed in all tissues examined except brain and heart. Highest levels in thymus, bone marrow and peripheral blood. -
Involvement in disease
Note=A chromosomal aberration involving RUNX1/AML1 is a cause of M2 type acute myeloid leukemia (AML-M2). Translocation t(8;21)(q22;q22) with RUNX1T1.
Note=A chromosomal aberration involving RUNX1/AML1 is a cause of therapy-related myelodysplastic syndrome (T-MDS). Translocation t(3;21)(q26;q22) with EAP or MECOM.
Note=A chromosomal aberration involving RUNX1/AML1 is a cause of chronic myelogenous leukemia (CML). Translocation t(3;21)(q26;q22) with EAP or MECOM.
Note=A chromosomal aberration involving RUNX1/AML1 is found in childhood acute lymphoblastic leukemia (ALL). Translocation t(12;21)(p13;q22) with TEL. The translocation fuses the 3'-end of TEL to the alternate 5'-exon of AML-1H.
Note=A chromosomal aberration involving RUNX1 is found in acute leukemia. Translocation t(11,21)(q13;q22) that forms a MACROD1-RUNX1 fusion protein.
Defects in RUNX1 are the cause of familial platelet disorder with associated myeloid malignancy (FPDMM) [MIM:601399]. FPDMM is an autosomal dominant disease characterized by qualitative and quantitative platelet defects, and propensity to develop acute myelogenous leukemia.
Note=A chromosomal aberration involving RUNX1/AML1 is found in therapy-related myeloid malignancies. Translocation t(16;21)(q24;q22) that forms a RUNX1-CBFA2T3 fusion protein.
Note=A chromosomal aberration involving RUNX1/AML1 is a cause of chronic myelomonocytic leukemia. Inversion inv(21)(q21;q22) with USP16. -
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. -
Post-translational
modificationsPhosphorylated in its C-terminus upon IL-6 treatment. Phosphorylation enhances interaction with MYST3.
Methylated. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 861 Human
- Omim: 151385 Human
- SwissProt: Q01196 Human
- Unigene: 149261 Human
- Unigene: 612648 Human
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Alternative names
- Acute myeloid leukemia 1 antibody
- Acute myeloid leukemia 1 protein antibody
- alpha subunit core binding factor antibody
see all
Images
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ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10^5 K-562 (Human chronic myelogenous leukemia lymphoblast) cells and 5µg of ab240639 [EPR23044-100]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
Additional screenshots of mapped reads can be downloaded here.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
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All lanes : Anti-RUNX1 / AML1 antibody [EPR23044-100] (ab240639) at 1/1000 dilution
Lane 1 : THP-1 (human monocytic leukemia monocyte), whole cell lysate
Lane 2 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate
Lane 3 : MOLT-4 (human lymphoblastic leukemia T lymphoblast), whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 48 kDa
Observed band size: 27-55 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 23352661, 29296779). The RUNX1 gene has several isoforms, 3 major isotypes. RUNX1b is broadly expressed, and RUNX1a overexpression has been reported in AML.
Exposure time: Lane 1: 48 seconds Lanes 2-3: 26 seconds
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RUNX1 / AML1 was immunoprecipitated from 0.35 mg Jurkat (human T cell leukemia T lymphocyte) whole cell lysate 10ug with ab240639 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab240639 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: Jurkat (human T cell leukemia T lymphocyte) whole cell lysate 10ug
Lane 2: ab240639 IP in Jurkat whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab240639 in Jurkat whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds
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Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling RUNX1 / AML1 with ab240639 at 1/2000 dilution (0.25 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human breast tissue is observed. The section was incubated with ab240639 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.
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Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling RUNX1 / AML1 with ab240639 at 1/2000 dilution (0.25 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human breast carcinoma (PMID: 24967588) tissue is observed. The section was incubated with ab240639 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.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Jurkat (Human T cell leukemia T lymphocyte) cells labelling RUNX1 / AML1 with ab240639 at 1/100 dilution, followed by ab240639 anti-RUNX1/AML1 ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing strong nuclear and weak cytoplasmic staining in Jurkat cell line is observed. Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab240639 anti-RUNX1/AML1 ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Jurkat (Human T cell leukemia T lymphocyte) cells labelling RUNX1 / AML1 with ab240639 at 1/500 (Red) compared with a Rabbit monoclonal IgG (ab172730, Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (1)
ab240639 has been referenced in 1 publication.
- Yuan C et al. A systematic dissection of the epigenomic heterogeneity of lung adenocarcinoma reveals two different subclasses with distinct prognosis and core regulatory networks. Genome Biol 22:156 (2021). PubMed: 34001209