ChIPAb+ CTCF

ChIPAb+ CTCF

Stock Code: 3586308
Manufacturer Part No: 17-10044
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Analysis Note


ControlIncludes negative control mouse IgG antibody and primers specific for human H19 ICR.


Application


The ChIPAb+ CTCF set includes the CTCF antibody, a negative control antibody (normal mouse IgG) & qPCR primers which amplify a 165 bp region of human H19 internal control region.


Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP with 15 min fixation) were subjected to chromatin immunoprecipitation using 2 µg of either Normal Mouse IgG or 2 µg of Anti-CTCF and the Magna ChIP™ G Kit (Cat. # 17-611).
Successful immunoprecipitation of CTCF associated DNA fragments was verified by qPCR using ChIP Primers, Human H19 ICR as a positive locus, and a representative published negative locus (Wendt, K.S., et al., 2008) (Please see figures). Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated.
Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.

Western Blot Analysis:
Representative lot data.
K562 cell lysate was resolved by electrophoresis, transferred to PVDF membranes and probed with Anti-CTCF, clone 1.2.1.5.4 (1 µg/mL).
Proteins were visualized using a Goat Anti-Mouse conjugated to HRP and a chemiluminescence detection system (Please see figures).

The following molecular weights have been reported in the literature: 180 kDa (CTCF-180), 130 kDa (CTCF-130), 82 kDa (CTCF-82), 70 kDa (CTCF-70). See Klenova, E.M., et al, (1997) Nucleic Acids Res. 25, 466–473 and Torrano,V., et al, (2006) Journal of Cell Science 119, 1746-1759) for additional details.
Please see molecular weight section for additional information.


General description


CTCF, also known as CCCTC-binding factor, contains 11 zinc finger (ZF) binding domains that allow it to bind different DNA target sequences and proteins. This multifunctional protein can act as a transcriptional activator or repressor. As a transcriptional repressor, CTCF, binds to promoters of vertebrate c-myc gene. It also binds to the PLK and PIM1 promoters. CTCF may prevent the access of transcriptional activators to enhancers. CTCF also acts as a transcriptional activator of APP. It is involved in different aspects of gene regulation including promoter activation or repression, hormone-responsive gene silencing, methylation-dependent chromatin insulation, and genomic imprinting. Mutations of CTCF in this gene have been associated with invasive breast cancers, prostate cancers, and Wilms tumors, suggesting a role for CTCF may as a tumor suppressor.


All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction.

Quality Level100
ManufacturerSIGMA-ALDRICH

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