CTCF polyclonal antibody - Classic | Antibody search engine

CTCF polyclonal antibody - Classic

Name	CTCF polyclonal antibody - Classic
Supplier	Diagenode
Catalog	C15410210
Prices	$295.00
Sizes	50 μg/ 20 μl
Host	Rabbit
Clonality	Polyclonal
Applications	ChIPseq ChIP-PCR WB ELISA
Species Reactivities	Human
Purity/Format	Affinity purified
Description	Rabbit Polyclonal
Gene	CTCF
Supplier Page	Shop

Product images

<strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against CTCF</strong><br />ChIP was performed with the Diagenode antibody against CTCF (Cat. No. C15410210) on sheared chromatin from 4,000,000 HeLa 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 optimized primers for the H19 imprinting control region, and a specific region in the GAPDH gene, used as positive controls, and for the Sat2 satellite repeat region, used as a negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Figure 1. ChIP results obtained with the Diagenode antibody directed against CTCF
ChIP was performed with the Diagenode antibody against CTCF (Cat. No. C15410210) on sheared chromatin from 4,000,000 HeLa 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 optimized primers for the H19 imprinting control region, and a specific region in the GAPDH gene, used as positive controls, and for the Sat2 satellite repeat region, used as a negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

<strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against CTCF</strong><br /> ChIP was performed on sheared chromatin from 4,000,000 HeLa cells using 1 μg of the Diagenode antibody against CTCF (Cat. No. C15410210) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq 2000. 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 3 mb region of the X-chromosome (figure 2A and B) and in two regions surrounding the H19 and GAPDH positive control genes, respectively (figure 2C, D and E). The position of the amplicon used for ChIP-qPCR is indicated by an arrow.

Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against CTCF
ChIP was performed on sheared chromatin from 4,000,000 HeLa cells using 1 μg of the Diagenode antibody against CTCF (Cat. No. C15410210) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq 2000. 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 3 mb region of the X-chromosome (figure 2A and B) and in two regions surrounding the H19 and GAPDH positive control genes, respectively (figure 2C, D and E). The position of the amplicon used for ChIP-qPCR is indicated by an arrow.

<strong> Figure 3. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody against CTCF (Cat. No. C15410210). 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 1:143,000.

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 against CTCF (Cat. No. C15410210). 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 1:143,000.

<strong>Figure 4. Western blot analysis using the Diagenode antibody directed against CTCF</strong><br /> Whole cell extracts (25 μg) from HeLa cells were analysed by Western blot using the Diagenode antibody against CTCF (Cat. No. C15410210) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.

Figure 4. Western blot analysis using the Diagenode antibody directed against CTCF
Whole cell extracts (25 μg) from HeLa cells were analysed by Western blot using the Diagenode antibody against CTCF (Cat. No. C15410210) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.