CTCF

CTCF CUTANA™ CUT&RUN Antibody

$495.00
SKU: 13-2014
Pack Size: 100 µL
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  • Type:  Monoclonal
  • Host:  Rabbit
  • Applications:  CUT&RUN, IHC, ICC, IP, WB
  • Reactivity:  Human, Mouse
  • Format:  Purified
  • Target Size: 83 kDa

Description

This antibody meets EpiCypher’s “CUTANA Compatible” criteria for performance in Cleavage Under Targets and Release Using Nuclease (CUT&RUN) and/or Cleavage Under Targets and Tagmentation (CUT&Tag) approaches to genomic mapping. Every lot of a CUTANA Compatible antibody is tested in the indicated approach using EpiCypher's optimized protocols and determined to yield peaks that show a genomic distribution pattern consistent with reported function(s) of the target protein. CTCF regulates transcription by binding DNA and preventing interaction between promoters and nearby enhancers or silencers.

Validation Data

CUT&RUN was performed on 500k K562 cells with either CTCF (0.125 µg) or IgG negative control (0.5 µg, EpiCypher 13-0042) antibodies using the CUTANA™ ChIC/CUT&RUN Kit v3.0 (EpiCypher 14-1048). Library preparation was performed with 5 ng of DNA (or the total amount recovered if less than 5 ng) using the CUTANA™ CUT&RUN Library Prep Kit (EpiCypher 14-1001/14-1002). Both kit protocols were adapted for high throughput Tecan liquid handling. Libraries were run on an Illumina NextSeq2000 with paired-end sequencing (2x50 bp). Sample sequencing depth was 7.6 million reads (IgG) and 10.4 million reads (CTCF). Data were aligned to the hg19 genome using Bowtie2. Data were filtered to remove duplicates, multi-aligned reads, and ENCODE DAC Exclusion List regions.

13-2014-ctcf-peaks (Click to enlarge)

Figure 1: CTCF peaks in CUT&RUN
CUT&RUN was performed as described above. Peaks were called with MACS2. Heatmaps show CTCF peaks relative to IgG negative control antibody in aligned rows ranked by intensity (top to bottom) and colored such that red indicates high localized enrichment and blue denotes background signal. CTCF showed expected enrichment around the TSS. Despite some observable background in the IgG control, differential enrichment with the target antibody is as expected.

13-2014-representative-browser-tracks (Click to enlarge)

Figure 2: CTCF CUT&RUN representative browser tracks
CUT&RUN was performed as described above. Gene browser shots were generated using the Integrative Genomics Viewer (IGV, Broad Institute). Three representative loci are shown.

13-2014-immunohistochemistry (Click to enlarge)

Figure 3: Immunohistochemistry data
FFPE sections of human colon carcinoma (left) and mouse spleen (right) using CTCF antibody at a dilution of 1:1,000.

13-2014-immunocytochemistry-data (Click to enlarge)

Figure 4: Immunocytochemistry data
FFPE section of human MCF-7 cells examined using CTCF antibody at a dilution of 1:1,000.

13-2014-immunoprecipitation-data (Click to enlarge)

Figure 5: Immunoprecipitation data
EpiCypher CTCF antibody (6 μg/mg lysate) was used to immunoprecipitate whole cell lysates (1 mg, 20% of IP loaded) isolated from HEK293T cells. A negative control IgG antibody and positive control antibodies (Bethyl Laboratories) were also used to demonstrate specificity of the IP. All CTCF antibodies target the same CTCF epitope, however, A300-534A-6 is hosted in rabbit. For blotting immunoprecipitates, EpiCypher CTCF antibody was used at a 1:1,000 dilution.

13-2014-western-blot-data (Click to enlarge)

Figure 6: Western blot data
Western analysis of CTCF in whole cell extracts from HEK293T, MCF-7, Jurkat, Hep-G2, and A-549 cells. Fifty micrograms of lysate was resolved via SDS-PAGE and detected with a 1:1,000 dilution of CTCF antibody.

Technical Information

Immunogen
Between amino acids 650 and 700
Storage
Stable for 1 year at 4°C from date of receipt
Formulation
Antigen affinity-purified recombinant monoclonal antibody in borate buffered saline pH 8.2, 0.1% BSA, 0.09% sodium azide

Recommended Dilution

CUT&RUN:
0.125 µg per reaction
Immunohistochemistry (IHC):
1:100 - 1:1,000. Epitope retrieval with citrate buffer pH 6.0 is recommended for FFPE tissue sections
Immunoprecipitation (IP):
20 µL/mg lysate
Western Blot (WB):
1:1,000

Gene & Protein Information

UniProt ID
P49711
Gene Name
CTCF
Protein Name
Transcriptional repressor CTCF
Target Size
83 kDa
Alternate Names
11-zinc finger protein, CCCTC-binding factor, CTCFL paralog

Documents & Resources

Technical Datasheet

Additional Info

Clone BLR041F is provided for commercial sale under license from Bethyl Laboratories with additional lot-specific validation in CUT&RUN performed by EpiCypher.

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