| Lot | A1329-001 |
|---|---|
| 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 | 1:500 | Fig 3 |
* Please note that the optimal antibody amount per ChIP should be determined by the end-user. We recommend testing 1-10 μl per IP.

Figure 1. ChIP results obtained with the Diagenode antibody directed against CBFb
ChIP assays were performed using SKNO-1 cells, the Diagenode antibody against CBFb (Cat. No. C15310002) 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, OGG1, NFE2, and SPI1 genes. Figure 1 shows the relative occupancy, calculated as the ratio + control/background for which the MYOG gene was used.




Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against CBFb
ChIP was performed as described above. The IP’d DNA from 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 region 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 human CBFb (Cat. No. C15310002). 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 1:8,800.
CBFb (UniProtKB/Swiss-Prot entry Q13951) represents the beta subunit of a heterodimeric core-binding transcription factor belonging to the PEBP2/CBF transcription factor family. These transcription factors regulate a host of genes specific to haematopoiesis (e.g. RUNX1) and osteogenesis (e.g. RUNX2). The beta subunit is the regulatory subunit which allosterically enhances the activity of the DNA binding alpha subunit as the complex binds to the core site of various enhancers and promoters. CBFb can be involved in a chromosomal rearrangement of chromosome 16 (inv(16)(p13q22)) which produces a fusion protein consisting of the N terminus of CBFb and the C-terminal portion of MYH11. This chromosomal rearrangement is associated with acute myeloid leukaemia of the M4Eo subtype.
| ELISA Enzyme-linked immunosorbent assay. Read more |
| ChIP-seq (ab) Read more |
| ChIP-qPCR (ab) Read more |
| Datasheet CBFb C15310002 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 |
How to properly cite our product/service in your workWe strongly recommend using this: CBFb Antibody (Hologic Diagenode Cat# C15310002 Lot# A1329-001). Click here to copy to clipboard. Using our products or services in your publication? Let us know! |
ERG and FLI1 binding sites demarcate targets for aberrant epigenetic regulation by AML1-ETO in acute myeloid leukemia. |