CTCF Recombinant Rabbit Monoclonal Antibody [JM10-61]
cat.: ET1703-90
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat, Zebrafish
Applications: WB, IF-Cell, IF-Tissue, IHC-P, IP, FC, CUT&Tag-seq, ChIP
Clonality: Monoclonal
Clone number: JM10-61
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 83 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human CTCF aa 685-727 / 727.
Positive control: HeLa cell lysate, 293T cell lysate, NIH/3T3 cell lysate, C6 cell lysate, PC-12 cell lysate, zebrafish tissue lysates, hybrid fish (crucian-carp) brain tissue lysates, HeLa, MCF-7, NIH/3T3, human liver tissue, human kidney tissue, mouse colon tissue, human endometrium tissue, rat colon tissue, rat kidney tissue.
Subcellular location: Nucleoplasm, Chromosome, centromere.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
  IP
1:1,000-1:2,000
1:100-1:500
1:100-1:500
1:50-1:2,000
1:500-1:1,000
Use at an assay dependent concentration.
Uniprot #: SwissProt: P49711 Human | Q61164 Mouse | Q9R1D1 Rat
Alternative names: 11 zinc finger protein 11 zinc finger transcriptional repressor 11-zinc finger protein CCCTC binding factor (zinc finger protein) CCCTC binding factor CCCTC-binding factor Ctcf CTCF_HUMAN CTCFL paralog MRD21 Transcriptional repressor CTCF
Images
ET1703-90_1.jpg Fig1: Western blot analysis of CTCF on different lysates with Rabbit anti-CTCF antibody (ET1703-90) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: 293T cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: C6 cell lysate
Lane 5: PC-12 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 83 kDa
Observed band size: 150 kDa

Exposure time: 4 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1703-90) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1703-90_2.jpg Fig2: Western blot analysis of CTCF on zebrafish tissue lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1703-90, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
ET1703-90_3.jpg Fig3: Western blot analysis of CTCF on hybrid fish (crucian-carp) brain tissue lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1703-90, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
ET1703-90_4.jpg Fig4: Immunocytochemistry analysis of HeLa cells labeling CTCF with Rabbit anti-CTCF antibody (ET1703-90) at 1/200 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-CTCF antibody (ET1703-90) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1703-90_5.jpg Fig5: ICC staining of CTCF in MCF-7 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1703-90, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1703-90_6.jpg Fig6: ICC staining of CTCF in NIH/3T3 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1703-90, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1703-90_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-CTCF antibody (ET1703-90) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-90) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1703-90_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-CTCF antibody (ET1703-90) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-90) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1703-90_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded mouse colon tissue using anti-CTCF antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-90, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1703-90_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded human endometrium tissue using anti-CTCF antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-90, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1703-90_11.jpg Fig11: Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-CTCF antibody (ET1703-90) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-90) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1703-90_12.jpg Fig12: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-CTCF antibody (ET1703-90) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-90) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1703-90_13.jpg Fig13: Flow cytometric analysis of HeLa cells labeling CTCF.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1703-90, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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