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17-10044 Sigma-AldrichChIPAb+ CTCF - ChIP Validated Antibody and Primer Set

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.

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Key Applications: Western Blotting, Dot Blot, Immunohistochemistry, Immunoprecipitation, Enzyme Immunoassay (ELISA), Multiplexing, Chromatin Immunoprecipitation (ChIP) Research Category: Epigenetics & Nuclear Function Research Sub-Category: Chromatin Biology, Transcription Factors

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Species Reactivity	Key Applications
H	WB, DB, IHC, IP, ELISA, Mplex, ChIP

Description
Catalogue Number	17-10044
Trade Name	ChIPAb+ Upstate
Description	ChIPAb+ CTCF - ChIP Validated Antibody and Primer Set
Overview	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. The ChIPAb+ CTCF set includes the CTCF antibody, a negative control antibody (normal mouse IgG), and qPCR primers which amplify a 165 bp region of human H19 internal control region. The CTCF and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of CTCF-associated chromatin.
Alternate Names	11 zinc finger transcriptional repressor 11-zinc finger protein CCCTC-binding factor CCCTC-binding factor (zinc finger protein) CTCFL paralog transcriptional repressor CTCF
Background Information	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.

References

Product Information
Format	Purified
Control	Includes negative control mouse IgG antibody and primers specific for human H19 ICR.
Presentation	Anti-CTCF, (mouse monoclonal IgG1κ. One vial containing 50 μg of protein G purified mouse monoclonal IgG1κ in buffer containing 0.1 M Tris-Glycine (pH 7.4, 150 mM NaCl) with 0.05% sodium azide, before the addition of glycerol to 30%. Store at -20°C. Normal Mouse IgG. Two vials containing 25 µg purified IgG in 25 µL storage buffer containing 0.1% sodium azide. Store at -20°C. ChIP Primers, Human H19 ICR. One vial containing 75 μL of 5 μM of each primer specific for human H19 internal control region. Store at -20°C. FOR: CCC ATC TTG CT GACC TCAC REV: AGA CCT GGG ACG TTT CTG TG
Quality Level	MQ100

Applications
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.
Key Applications	Western Blotting Dot Blot Immunohistochemistry Immunoprecipitation Enzyme Immunoassay (ELISA) Multiplexing Chromatin Immunoprecipitation (ChIP)
Application Notes	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.

Biological Information
Immunogen	His-tagged recombinant protein corresponding to human CTCF.
Clone	1.2.1.5.4
Host	Mouse
Specificity	Recognizes CTCF, Mr ~130 kDa.
Isotype	IgG1κ
Species Reactivity	Human
Antibody Type	Monoclonal Antibody
Entrez Gene Number	NP_006556.1
Entrez Gene Summary	This gene is a member of the BORIS + CTCF gene family and encodes a transcriptional regulator protein with 11 highly conserved zinc finger (ZF) domains. This nuclear protein is able to use different combinations of the ZF domains to bind different DNA target sequences and proteins. Depending upon the context of the site, the protein can bind a histone acetyltransferase (HAT)-containing complex and function as a transcriptional activator or bind a histone deacetylase (HDAC)-containing complex and function as a transcriptional repressor. If the protein is bound to a transcriptional insulator element, it can block communication between enhancers and upstream promoters, thereby regulating imprinted expression. Mutations in this gene have been associated with invasive breast cancers, prostate cancers, and Wilms' tumors. [provided by RefSeq].
Gene Symbol	CTCF
Purification Method	Protein G Purified
UniProt Number	P49711
UniProt Summary	Function: Transcriptional repressor binding to promoters of vertebrate c-myc gene. Also binds to the PLK and PIM1 promoters. May prevent the access of transcriptional activators to enhancers. Also acts as a transcriptional activator of APP. Involved in different aspects of gene regulation including promoter activation or repression, hormone-responsive gene silencing, methylation- dependent chromatin insulation, and genomic imprinting. Seems to act as tumor suppressor.SIZE: 727 amino acids; 82785 Da Subcellular Location Nucleus. Tissue Specificity:Ubiquitous. Absent in primary spermatocytes. Similarity: Belongs to the CTCF zinc-finger protein family. & Contains 11 C2H2-type zinc fingers.
Molecular Weight	~ 130 kDa Note: Poly(ADP-ribosyl)ated isoform, C-terminal truncation, or sumoylation will cause variable migration of CTCF protein on SDS–PAGE, which may present CTCF with different molecular weight on Western Blot. Modifications are dependent on cell type and growing condition. 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.

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Materials Information

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Safety Information according to GHS

Safety Information

Product Usage Statements
Quality Assurance	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 (Please see figures). Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
Usage Statement	Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Product Usage Statements

Quality Assurance

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 (Please see figures).
Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.

Usage Statement

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage and Shipping Information
Storage Conditions	Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.

Storage and Shipping Information

Storage Conditions

Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.

Packaging Information
Material Size	25 assays
Material Package	25 assays per set. Recommended use: ~2 μg of antibody per chromatin immunoprecipitation (dependent upon biological context).

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Supplemental Information

Specifications

Global Trade Item Number
Catalog Number	GTIN
17-10044	04053252679230

Documentation

ChIPAb+ CTCF - ChIP Validated Antibody and Primer Set SDS

Title
Safety Data Sheet (SDS)

ChIPAb+ CTCF - ChIP Validated Antibody and Primer Set Certificates of Analysis

Title	Lot Number
ChIPAb+ CTCF - 2108581	2108581
ChIPAb+ CTCF - 2377389	2377389
ChIPAb+ CTCF - 2433563	2433563
ChIPAb+ CTCF - 2006937	2006937
ChIPAb+ CTCF - 2246838	2246838
ChIPAb+ CTCF - 3150649	3150649
ChIPAb+ CTCF - 3380454	3380454
ChIPAb+ CTCF - 3529581	3529581
ChIPAb+ CTCF - 3562370	3562370
ChIPAb+ CTCF - 3698765	3698765

Brochure

Title
Advance your Epigenetics Research

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