ETO Antibody - ChIP-seq Grade (C15310001)

Lot	A710-001
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	1:100	Fig 3

ETO Antibody ChIP Grade

Figure 1. ChIP results obtained with the Diagenode antibody directed against ETO
ChIP assays were performed using SKNO-1 cells, the Diagenode antibody against ETO (Cat. No. C15310001) 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, OGG1 and VEGF genes. Figure 1 shows the occupancy, calculated as the ratio + control/background for which the H2B gene was used.

ETO Antibody ChIP-seq Grade

ETO Antibody for ChIP-seq

ETO Antibody for ChIP-seq assay

ETO Antibody validated in ChIP-seq

Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against 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.

ETO Antibody ELISA Validation

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 human ETO (Cat. No. C15310001). 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 1:1,300.

Target Description

ETO (UniProtKB/Swiss-Prot entry Q06455) is a transcriptional regulator which belongs to the myeloid translocation gene family. ETO exerts its function by interaction with transcription factors bound to promoters and binding to histone deacetylases. It recruits a range of corepressors to facilitate transcriptional repression. The t(8;21)(q22;q22) translocation is one of the most frequent karyotypic abnormalities in acute myeloid leukaemia. This translocation produces a chimeric gene made up of the 5’-region of AML1and the 3’-region of the ETO gene. The chimeric protein is thought to associate with the nuclear corepressor/ histone deacetylase complex to block hematopoietic differentiation.

Applications

ELISA
Enzyme-linked immunosorbent assay. Read more

ChIP-seq (ab)
Read more

ChIP-qPCR (ab)
Read more

Documents

Datasheet ETO C15310001 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

Publications

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ERG and FLI1 binding sites demarcate targets for aberrant epigenetic regulation by AML1-ETO in acute myeloid leukemia.
Martens JH, Mandoli A, Simmer F, Wierenga BJ, Saeed S, Singh AA, Altucci L, Vellenga E, Stunnenberg HG
ERG and FLI1 are closely related members of the ETS family of transcription factors and have been identified as essential factors for the function and maintenance of normal hematopoietic stem cells. Here, genome-wide analysis revealed that both ERG and FLI1 occupy similar genomic regions as AML1-ETO in t(8;21) AMLs ...

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ETO Antibody - ChIP-seq Grade

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