CTCF polyclonal antibody

Lot	A2359-00234P
Concentration	1.5 µg/µl
Species reactivity	Human, mouse
Type	Polyclonal

Purity	Affinity purified
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 and 0.05% ProClin 300.
Precautions	This product is for research use only. Not for use in diagnostic or therapeutic procedures.

Applications	Suggested dilution	References
ChIP ^*	1 µg per ChIP	Fig 1, 2
ELISA	1:10,000	Fig 3
Western Blotting	1:1,000	Fig 4, 5
Immunofluorescence	1:200	Fig 6

ChIP

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 MYOD1 gene and 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).

ChIP-seq figure A

ChIP-seq figure B

ChIP-seq figure C

ChIP-seq figure D

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 1 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 and D).

ELISA

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 >1:500,000.

Western blot

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.

Western blot

Figure 5. Western blot analysis using the Diagenode antibody directed against CTCF
Whole cell extracts (40 μg) from HeLa cells transfected with CTCF siRNA (lane 2) and from an untransfected control (lane 1) 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.

Immunofluorescence

Figure 6. Immunofluorescence using the Diagenode antibody directed against CTCF
HeLa cells were stained with the Diagenode antibody against CTCF (Cat. No. C15410210) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the CTCF antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

お客様の声

In life sciences, epigenetics is nowadays the most rapid developing field with new astonishing discoveries made every day. To keep pace with this field, we are in need of reliable tools to foster our research - tools Diagenode provides us with. From antibodies to automated solutions - all from one source and with robust support. Antibodies used in our lab: H3K27me3 polyclonal antibody – Premium, H3K4me3 polyclonal antibody – Premium, H3K9me3 polyclonal antibody – Premium, H3K4me1 polyclonal antibody – Premium, CTCF polyclonal antibody – Classic, Rabbit IgG.
Dr. Florian Uhle, Dept. of Anesthesiology, Heidelberg University Hospital, Germany

実験手法

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

ELISA
Enzyme-linked immunosorbent assay. Read more

ChIP-seq (ab)
Read more

ChIP-qPCR (ab)
Read more

IF
Immunofluorescence: Diagenode offers huge selection of highly sensitive antibodies validated in IF. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9 HeLa cells transfected with a Cas9 expression vector (... Read more

siRNA Knockdown
Epigenetic antibodies you can trust! Antibody quality is essential for assay success. Diagenode offers antibodies that are actually validated and have been widely used and published by the scientific community. Now we are adding a new level o... Read more

資料

Datasheet CTCF C15410210 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

出版物

How to properly cite this product in your work

Diagenode strongly recommends using this: CTCF polyclonal antibody - Classic (sample size) (Diagenode Cat# C15410210-10 Lot# A2359-00234P). Click here to copy to clipboard.

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Preformed chromatin topology assists transcriptional robustness of during limb development.
Paliou C, Guckelberger P, Schöpflin R, Heinrich V, Esposito A, Chiariello AM, Bianco S, Annunziatella C, Helmuth J, Haas S, Jerković I, Brieske N, Wittler L, Timmermann B, Nicodemi M, Vingron M, Mundlos S, Andrey G
Long-range gene regulation involves physical proximity between enhancers and promoters to generate precise patterns of gene expression in space and time. However, in some cases, proximity coincides with gene activation, whereas, in others, preformed topologies already exist before activation. In this study, we inves...

High-throughput ChIPmentation: freely scalable, single day ChIPseq data generation from very low cell-numbers.
Gustafsson C, De Paepe A, Schmidl C, Månsson R
BACKGROUND: Chromatin immunoprecipitation coupled to sequencing (ChIP-seq) is widely used to map histone modifications and transcription factor binding on a genome-wide level. RESULTS: We present high-throughput ChIPmentation (HT-ChIPmentation) that eliminates the need for DNA purification prior to library amplifica...

DNA methylation signatures follow preformed chromatin compartments in cardiac myocytes
Nothjunge S. et al.
Storage of chromatin in restricted nuclear space requires dense packing while ensuring DNA accessibility. Thus, different layers of chromatin organization and epigenetic control mechanisms exist. Genome-wide chromatin interaction maps revealed large interaction domains (TADs) and higher order A and B compartments, r...

High Resolution Mapping of Chromatin Conformation in Cardiac Myocytes Reveals Structural Remodeling of the Epigenome in Heart Failure
Rosa-Garrido M. et al.
Background -Cardiovascular disease is associated with epigenomic changes in the heart, however the endogenous structure of cardiac myocyte chromatin has never been determined. Methods -To investigate the mechanisms of epigenomic function in the heart, genome-wide chromatin conformation capture (Hi-C) and DNA sequenc...

Platelet function is modified by common sequence variation in megakaryocyte super enhancers
Petersen R. et al.
Linking non-coding genetic variants associated with the risk of diseases or disease-relevant traits to target genes is a crucial step to realize GWAS potential in the introduction of precision medicine. Here we set out to determine the mechanisms underpinning variant association with platelet quantitative traits usi...

Loss of cohesin complex components STAG2 or STAG3 confers resistance to BRAF inhibition in melanoma
Shen CH et al.
The protein kinase B-Raf proto-oncogene, serine/threonine kinase (BRAF) is an oncogenic driver and therapeutic target in melanoma. Inhibitors of BRAF (BRAFi) have shown high response rates and extended survival in patients with melanoma who bear tumors that express mutations encoding BRAF proteins mutant at Val600, ...

CTCF polyclonal antibody - Classic (sample size)

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CTCF polyclonal antibody - Classic (sample size)

How to properly cite this product in your work

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CTCF polyclonal antibody - Classic

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