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Polyclonal antibody raised in rabbit against human EHF (ETS Homologous Factor), using two synthetic peptides containing a sequence from the central part and the C-terminus of the protein, respectively.
Lot
A2883P
Concentration
0.8 µg/µl
Species reactivity
Human: positive. Other species: not tested.
Type
Polyclonal
Purity
Affinity purified polyclonal antibody.
Host
Rabbit
Storage Conditions
Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles.
Storage Buffer
PBS containing 0.05% azide.
Precautions
This product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications
Suggested dilution
References
ChIP/ChIP-seq*
1 – 2 µg per ChIP
Fig 1, 2
ELISA
1:1,000 - 1:50,000
Fig 3
Western Blotting
Not recommended
*Please note that the optimal antibody amount per ChIP should be determined by the end-user. We recommend testing 1-5 µg per ChIP.
Figure 1. ChIP results obtained with the Diagenode antibody directed against EHF ChIP assays were performed using T47D cells, the Diagenode antibody against EHF (Cat. No. C15410363) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010055), using sheared chromatin from 4 million cells. A titration consisting of 1, 2, 5 and 10 µg of antibody per ChIP experiment was analyzed. IgG (2 µg/IP) was used as a negative IP control. Quantitative PCR was performed with primers for the promoters of the ITGA2 and OAS3 genes, used as positive controls, and for the MYOD1 gene and the Sat2 satellite repeat, used as negative controls.
A.
B.
C.
D.
Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against EHF ChIP was performed on sheared chromatin from 4,000,000 T47D cells using 2 µg of the Diagenode antibody against EHF (Cat. No. C15410363) as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq 4000. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution along the complete sequence and a 300 kb region of chromosome 1 (figure 2A and B) and in two genomic regions surrounding the ITGA2 and OAS3 positive control genes, respectively (figure 2C and D).
Figure 3. Determination of the antibody titer To determine the titer of the antibody, an ELISA was performed using a serial dilution of Diagenode antibody directed against EHF (Cat. No. C15410363). 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 1:98,000.
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