Anti-CTCF antibody [RM1044]

7 product images

• 

  Western blot - Anti-CTCF antibody [RM1044] (ab300639)

  CTCF Western blot staining using rabbit Anti-CTCF antibody

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  The multiples bands observed in the WB have been reported in literature (PMID: 12878173, PMID: 8246978).

  All lanes: Western blot - Anti-CTCF antibody [RM1044] (ab300639) at 1/1000 dilution

  Lane 1: Human spleen lysate at 20 µg

  Lane 2: Mouse heart lysate at 20 µg

  Lane 3: Mouse spleen lysate at 20 µg

  Lane 4: Mouse colon lysate at 20 µg

  Lane 5: Rat heart lysate at 20 µg

  Lane 6: Rat spleen lysate at 20 µg

  Lane 7: Rat colon lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Predicted band size: 83 kDa

  Observed band size: 140 kDa, 130 kDa, 97 kDa, 80 kDa, 73 kDa, 55 kDa, 45 kDa

• 

  Immunoprecipitation - Anti-CTCF antibody [RM1044] (ab300639)

  CTCF was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab300639 at 1/30 dilution (2µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab300639 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

  Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  All lanes: Immunoprecipitation - Anti-CTCF antibody [RM1044] (ab300639) at 1/30 dilution

  All lanes: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate

  Secondary

  All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

  Predicted band size: 83 kDa

  Observed band size: 40 kDa, 130 kDa, 97 kDa, 80 kDa, 73 kDa, 55 kDa, 45 kDa

  Exposure time: 10s

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTCF antibody [RM1044] (ab300639)

  Immunohistochemical analysis of paraffin-embedded human breast tissue labelling CTCF with ab300639 at 1/10000 (0.053 µg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Nuclear staining on human breast is observed. The section was incubated with ab300639 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

  Secondary antibody only control: PBS was used instead of primary antibody followed by ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTCF antibody [RM1044] (ab300639)

  Immunohistochemical analysis of paraffin-embedded rat colon tissue labelling CTCF with ab300639 at 1/10000 (0.053 µg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Nuclear staining on rat colon. The section was incubated with ab300639 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

  Secondary antibody only control: PBS was used instead of primary antibody followed by ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins was used.

• 

  Flow Cytometry (Intracellular) - Anti-CTCF antibody [RM1044] (ab300639)

  Intracellular flow cytometric analysis of 4% paraformaldehyde fixed and 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling CTCF with ab300639 at 1/500 dilution (0.1µg) (red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-CTCF antibody [RM1044] (ab300639)

  Immunofluorescent analysis of 4% paraformaldehyde fixed and 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) labeling CTCF with ab300639 at 1/2000 dilution (0.266 μg/ml), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488, green) preadsorbed at 1:1000 (2 μg/mL). Confocal image showing nuclear staining in HeLa cell line.

  Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) (red) was used as a counterstain at 1:200 dilution (2.5 μg/ml). Nuclear counter satin is DAPI.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CTCF antibody [RM1044] (ab300639)

  Immunohistochemical analysis of paraffin-embedded mouse colon tissue labelling CTCF with ab300639 at 1/10000 (0.053 µg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Nuclear staining on mouse colon. The section was incubated with ab300639 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

  Secondary antibody only control: PBS was used instead of primary antibody followed by ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins was used.