Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23309-113] to RUNX1 / AML1 - ChIP Grade
- Suitable for: Flow Cyt, WB, ChIP, IP
- Reacts with: Human
Product nameAnti-RUNX1 / AML1 antibody [EPR23309-113] - ChIP Grade
See all RUNX1 / AML1 primary antibodies
DescriptionRabbit monoclonal [EPR23309-113] to RUNX1 / AML1 - ChIP Grade
Tested applicationsSuitable for: Flow Cyt, WB, ChIP, IPmore details
Unsuitable for: ICC/IF or IHC-P
Species reactivityReacts with: Human
Recombinant fragment within Human RUNX1/ AML1 aa 50-200. The exact sequence is proprietary.
Database link: Q01196
- WB: Jurkat, MOLT-4 and THP-1 whole cell lysates. Flow Cyt: Jurkat cells. IP: Jurkat and K562 whole cell lysate.ChIP: Chromatin prepared from K562 cells.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Storage instructionsShipped 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 bufferPreservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab272456 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000. Detects a band of approximately 27-48 kDa (predicted molecular weight: 48 kDa).|
|ChIP||Use 5 µg for 25 µg of chromatin.|
FunctionCBF 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 specificityExpressed in all tissues examined except brain and heart. Highest levels in thymus, bone marrow and peripheral blood.
Involvement in diseaseNote=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 similaritiesContains 1 Runt domain.
DomainA proline/serine/threonine rich region at the C-terminus is necessary for transcriptional activation of target genes.
modificationsPhosphorylated in its C-terminus upon IL-6 treatment. Phosphorylation enhances interaction with MYST3.
- Information by UniProt
- Acute myeloid leukemia 1 antibody
- Acute myeloid leukemia 1 protein antibody
- alpha subunit core binding factor antibody
Chromatin was prepared from K-562 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 ab272456 (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 from paper PMID：20959602
The RUNX1 enrichment profile is consistent with what have been described in literature (PMID: 20959602).
All lanes : Anti-RUNX1 / AML1 antibody [EPR23309-113] - ChIP Grade (ab272456) at 1/1000 dilution
Lane 1 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 2 : MOLT-4 (human lymphoblastic leukemia t lymphoblast) whole cell lysate
Lane 3 : THP-1 (human monocytic leukemia monocyte) whole cell lysate
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution
Predicted band size: 48 kDa
Observed band size: 27-48 kDa why is the actual band size different from the predicted?
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
RUNX1 has several isoforms. The molecular weight observed is consistent with what have been described in literature (PMID:17431130, 29296779).
Exposure time: 3 minutes
RUNX1 / AML1 was immunoprecipitated from 0.35 mg Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate with ab272456 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab272456 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate 20 ug
Lane 2: ab272456 IP in Jurkat whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab272456 in Jurkat whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 32 seconds.
Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized Jurkat (Human T cell leukemia T lymphocyte) cells labelling RUNX1 / AML1 with ab272456 at 1/500 dilution (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.
RUNX1 / AML1 was immunoprecipitated from K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate with ab272456 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab272456 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: K562 whole cell lysate 10 µg
Lane 2: ab272456 IP in K562 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab272456 in K562 whole cell lysate
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 3 mins.
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.
ab272456 has not yet been referenced specifically in any publications.