CTCF

CTCF CUTANA™ CUT&RUN Antibody

$445.00
SKU: 13-2014
Pack Size: 100 µL
  • Type:  Monoclonal [BLR041F]
  • Host:  Rabbit
  • Target Size:  83 kDa
  • Format:  Purified IgG
  • Reactivity:  Human, Mouse
  • Applications: CUT&RUN, WB, IP, IHC, ICC

Description

This CTCF 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 CUTANA approach using EpiCypher optimized protocols and determined to yield peaks that show a genomic distribution pattern consistent with reported function(s) of the target protein. CTCF antibody produces CUT&RUN peaks at promoters (Figure 1A) and to a greater extent within intergenic and intronic regions (Figure 1B). CUT&RUN peaks overlap with known CTCF transcription factor binding motifs (Figure 2).

Validation Data

Figure 1: CTCF signal enrichment in CUT&RUN
CUT&RUN was performed using 500,000 K562 cells with CTCF antibody (0.125 µg) and IgG control (0.5 µg, EpiCypher 13-0042). (A) Sequencing reads were aligned to annotated TSSs (+/- 2 kbp) of 18,793 genes. High, medium, and low signal is ranked by intensity (top to bottom; aligned relative to CTCF) and reflected by red, yellow, and blue colors, respectively. (B) CTCF peaks were called using SEACR. The number of CTCF peaks which fall into distinct classes of functionally annotated genomic regions is plotted.

Figure 2: CTCF transcription factor binding motif analysis in CUT&RUN
(A) MEME analysis determined that the CTCF consensus motif, represented as a sequence logo position weight matrix, was enriched under CUT&RUN peaks produced by CTCF antibody. (B) The number of CTCF peaks containing consensus motifs from panel A is represented by a Venn Diagram. (C) Three representative loci showing CTCF peaks (IGV).

Figure 3: Western blot detection of human and mouse CTCF
Whole cell lysates were isolated from HEK293T (293T), MCF-7, Hep-G2, A-549, SW620, SK-MEL-28 (SK), Jurkat, TCMK-1, and NIH 3T3 cells using NETN lysis buffer. Fifty micrograms (50 µg) of each lysate was loaded onto an SDS-PAGE gel and analyzed under standard western blot conditions using CTCF antibody (1;1,000).

Figure 4: Immunoprecipitation of human CTCF
EpiCypher CTCF antibody (20 µL) was used to immunoprecipitate whole cell lysates isolated from HEK293T cells using NETN lysis buffer (1 mg per IP). A negative control IgG antibody and a different CTCF positive control antibody (Bethyl Laboratories) were also used to demonstrate specificity of the IP. Immunoprecipitates were loaded onto an SDS-PAGE gel (20% of IP loaded) and probed via western blot with EpiCypher CTCF antibody (1:1,000).

Figure 5: Immunohistochemistry detection of human CTCF
FFPE section of human lung carcinoma examined using CTCF antibody (1:100 dilution).

Figure 6: Immunocytochemistry detection of human CTCF
FFPE section of human MCF-7 cells examined using CTCF antibody (1:100 dilution).

Technical Information

Immunogen
A synthetic peptide corresponding to human CTCF (CCCTC-binding factor) amino acids 650 to 700.
Formulation
Purified recombinant monoclonal CTCF antibody (100 µg/mL) in borate buffered saline pH 8.2, 0.1% BSA, 0.09% sodium azide.
Storage and Stability
Stable for 1 year at 4°C from date of receipt.

Application Notes

Recommended Dilutions:

CUT&RUN: 0.1 - 0.3 µg

Western Blot (WB): 1:1,000

Immunoprecipitation (IP): 20 µL/mg lysate

Immunohistochemistry (IHC): 1:100 - 1:500*

Immunocytochemistry (ICC): 1:100 - 1:500*

*Epitope retrieval with citrate buffer pH 6.0 recommended for FFPE tissues.

Documents & Resources

Additional Info

This CTCF antibody is provided for commercial sale under license from Bethyl Laboratories, Inc.

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