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CTCF (D31H2) XP® Rabbit mAb
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Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the carboxy terminus of the human CTCF protein.
Product Usage Information
For optimal ChIP or ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
The CUT&Tag dilution was determined using CUT&Tag Assay Kit #77552.
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:50 |
| Immunohistochemistry (Paraffin) | 1:640 - 1:2560 |
| Immunofluorescence (Frozen) | 1:200 - 1:800 |
| Immunofluorescence (Immunocytochemistry) | 1:400 - 1:1600 |
| Chromatin IP | 1:50 |
| Chromatin IP-seq | 1:50 |
| CUT&RUN | 1:50 |
| CUT&Tag | 1:50 |
Specificity/Sensitivity
Species Reactivity:
Human, Mouse, Rat, Monkey
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
For a carrier free (BSA and azide free) version of this product see product #48859.
参考图片
CUT&Tag was performed with HCT 116 cells and CTCF (D31H2) XP® Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figures show binding across chromosome 8 (upper), including MYC (lower), a known target gene of CTCF (see our ChIP-qPCR figure).
Western blot analysis of extracts from various cell lines using CTCF (D31H2) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using CTCF (D31H2) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using CTCF (D31H2) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human lymphoma using CTCF (D31H2) XP® Rabbit mAb.
Confocal immunofluorescent analysis of HCT-116 cells using CTCF (D31H2) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
Chromatin immunoprecipitations were performed with cross-linked chromatin from Hela cells and CTCF (D31H2) XP® Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across H19, a known target gene of CTCF (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.
Chromatin immunoprecipitations were performed with cross-linked chromatin from Hela cells and CTCF (D31H2) XP® Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 11 (upper), including H19 (lower), a known target gene of CTCF (see additional figure containing ChIP-qPCR data).
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells, with either CTCF (D31H2) XP® Rabbit mAb or Normal Rabbit IgG #2729, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human c-Myc promoter primers, SimpleChIP® Human H19/Igf2 ICR Primers #5172, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
CUT&RUN was performed with HCT 116 cells and CTCF (D31H2) XP® Rabbit mAb #3418, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across MYC, a known target gene of CTCF (see additional figure containing CUT&RUN-qPCR data).
CUT&RUN was performed with HCT 116 cells and CTCF (D31H2) XP® Rabbit mAb #3418, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 8 (upper), including MYC (lower), a known target gene of CTCF (see additional figure containing CUT&RUN-qPCR data).
CUT&RUN was performed with HCT 116 cells and CTCF (D31H2) XP® Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT & RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using human c-Myc promoter primers, SimpleChIP® Human H19/Igf2 ICR Primers #5172, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
CUT&Tag was performed with HCT 116 cells and CTCF (D31H2) XP® Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figure shows binding across MYC, a known target gene of CTCF (see our ChIP-qPCR figure).