AML1-ETO Antibody
Alternative name: RUNX1-RUNX1T1
Polyclonal antibody raised in rabbit against the AML1-ETO fusion protein using a KLH-conjugated synthetic peptide.
| Lot | A706-002 |
|---|---|
| Concentration | not determined |
| Species reactivity | Human |
| Type | Polyclonal |
| Purity | Whole antiserum |
| Host | Rabbit |
| Precautions | This product is for research use only. Not for use in diagnostic or therapeutic procedures. |
| Applications | Suggested dilution | References |
|---|---|---|
| ChIP/ChIP-seq * | 4 μl/ChIP | Fig 1, 2 |
| ELISA | 1:500 | Fig 3 |
| Western Blotting | 1:1,000 | Fig 4 |
* Please note that the optimal antibody amount per ChIP should be determined by the end-user. We recommend testing 1-10 μl per IP.
Validation Data
Figure 1. ChIP results obtained with the Diagenode antibody directed against AML1-ETO
ChIP assays were performed using Kasumi-1 cells, the Diagenode antibody against AML1-ETO (Cat. No. C15310197) and optimized primer pairs for qPCR. Sheared chromatin from 1.25 million cells and 4 μl of antibody were used per ChIP experiment. QPCR was performed using primers specific for the FUT7, NFE2 and OGG1 genes. Figure 1 shows the occupancy, calculated as the ratio + control/background for which the promoter of the H2B gene was used.
A.ChIP-seq signals on chromosome 3
B.Genomic region on chromosome 3 surrounding the OGG1 gene
C. Genomic region on chromosome 9 surrounding the FUT7 gene
D. Genomic region on chromosome 12 surrounding the NFE2 gene
Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against AML1-ETO
ChIP was performed as described above. The IP’d DNA of 6 ChIP’s was pooled and analysed with an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 32 bp tags were aligned to the human reference genome (hg18) using the ELAND algorithm. Figure 2 shows the results of the complete chromosome 3 and three genomic regions surrounding the OGG1, FUT7 and NFE2 genes, respectively. The position of the PCR amplicon is indicated with an arrow.
Figure 3. Determination of the antibody titer
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against AML1-ETO (Cat. No. C15310197). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:32,750.
Figure 4. Western blot analysis using the Diagenode antibody directed against AML1-ETO
Nuclear extracts of SKNO-1 cells (15 μg) were analysed by Western blot using the Diagenode antibody against AML1-ETO (Cat. No. C15310197) diluted 1:1,000 in TBSTween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.
Target Description
This antibody specifically recognizes the AML1 (RUNX1) (UniProtKB/Swiss-Prot entry Q01196) - ETO (RUNX1T1) (UniProtKB/ Swiss-Prot entry Q06455) fusion protein that arises due to a translocation between chromosome 8 and 22 (t(8;21)(q22;q22)). This translocation is one of the most frequent karyotypic abnormalities observed in acute myeloid leukaemia. It produces a chimerical gene made up of the 5’-region of AML1and the 3’-region of ETO. The chimerical protein is thought to associate with the nuclear corepressor/histone deacetylase complex to block hematopoietic differentiation.
Applications
| ELISA Enzyme-linked immunosorbent assay. Read more |
| WB Western blot : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies. Learn more about: Load... Read more |
| ChIP-seq (ab) Read more |
| ChIP-qPCR (ab) Read more |
Documents
| Datasheet AML1-ETO C15310197 DATASHEET Datasheet description | Download |
| Antibodies you can trust POSTER Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar... | Download |
| Epigenetic Antibodies Brochure BROCHURE More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e... | Download |
Safety sheets
Publications
How to properly cite our product/service in your workWe strongly recommend using this: AML1-ETO Antibody (Hologic Diagenode Cat# C15310197 Lot# A706-002). Click here to copy to clipboard. Using our products or services in your publication? Let us know! |
RUNX1-ETO Depletion in t(8;21) AML Leads to C/EBPα- and AP-1-Mediated Alterations in Enhancer-Promoter Interaction. |
Suppression of RUNX1/ETO oncogenic activity by a small molecule inhibitor of tetramerization |
The Hematopoietic Transcription Factors RUNX1 and ERG Prevent AML1-ETO Oncogene Overexpression and Onset of the Apoptosis Program in t(8;21) AMLs |
MEIS2 Is an Oncogenic Partner in AML1-ETO-Positive AML |
Loss of TET2 in hematopoietic cells leads to DNA hypermethylation of active enhancers and induction of leukemogenesis |
Immune evasion by oncogenic proteins of acute myeloid leukemia. |
ERG and FLI1 binding sites demarcate targets for aberrant epigenetic regulation by AML1-ETO in acute myeloid leukemia. |