Anti-CNTF antibody [EPR24553-104]

12 product images

• 

  Western blot - Anti-CNTF antibody [EPR24553-104] (ab270992)

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

  Bands at 37, 50KD are likely to be multimer forms of CNTF (PMID: 9236223).

  Negative control: Brain, stomach (PMID: 12855189).

  Exposure time: 59 seconds.

  All lanes: Western blot - Anti-CNTF antibody [EPR24553-104] (ab270992) at 1/1000 dilution

  Lane 1: Mouse sciatic nerve lysate at 20 µg

  Lane 2: Mouse brain lysate at 20 µg

  Lane 3: Mouse stomach 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 1/10000 dilution

  Predicted band size: 23 kDa

  Observed band size: 22 kDa, 37 kDa, 50 kDa

• 

  Western blot - Anti-CNTF antibody [EPR24553-104] (ab270992)

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

  Negative control: Stomach (PMID: 12855189).

  Exposure time: 81 seconds.

  All lanes: Western blot - Anti-CNTF antibody [EPR24553-104] (ab270992) at 1/1000 dilution

  Lane 1: Rat sciatic nerve lysate at 20 µg

  Lane 2: Rat spinal cord lysate at 20 µg

  Lane 3: Rat stomach 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 1/10000 dilution

  Predicted band size: 23 kDa

  Observed band size: 22 kDa

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CNTF antibody [EPR24553-104] (ab270992)

  Immunohistochemical analysis of paraffin-embedded Mouse sciatic nerve tissue labelling CNTF with ab270992 at 1/500 dilution (0.962 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining in mouse sciatic nerve. The section was incubated with ab270992 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
  

  Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CNTF antibody [EPR24553-104] (ab270992)

  Immunohistochemical analysis of paraffin-embedded Rat sciatic nerve tissue labelling CNTF with ab270992 at 1/500 dilution (0.962 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining in rat sciatic nerve. The section was incubated with ab270992 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
  

  Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CNTF antibody [EPR24553-104] (ab270992)

  Immunohistochemical analysis of paraffin-embedded Mouse stomach tissue labelling CNTF with ab270992 at 1/500 dilution (0.962 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control: No staining in mouse stomach. The section was incubated with ab270992 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
  

  Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• 

  Immunohistochemistry (Frozen sections) - Anti-CNTF antibody [EPR24553-104] (ab270992)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse sciatic nerve tissue labeling CNTF with ab270992 at 1/100 dilution (4.81 ug/ml) followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on mouse sciatic nerve is observed. The nuclear counterstain was DAPI (Blue).
  

  Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• 

  Immunohistochemistry (Frozen sections) - Anti-CNTF antibody [EPR24553-104] (ab270992)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse stomach tissue labeling CNTF with ab270992 at 1/100 dilution (4.81 ug/ml) followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Negative control: No staining on mouse stomach (PMID: 12855189) is observed. The nuclear counterstain was DAPI (Blue).
  

  Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• 

  Immunoprecipitation - Anti-CNTF antibody [EPR24553-104] (ab270992)

  CNTF was immunoprecipitated from 0.35 mg Mouse sciatic nerve lysate with ab270992 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab270992 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
  

  Lane 1: Mouse sciatic nerve lysate 10ug

  Lane 2: ab270992 IP in Mouse sciatic nerve lysate

  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab270992 in mouse sciatic nerve lysate

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

  Exposure time: 10 seconds.

  Bands at 37, 50KD are likely to be multimer forms of CNTF (PMID: 9236223).

  All lanes: Immunoprecipitation - Anti-CNTF antibody [EPR24553-104] (ab270992)

  Predicted band size: 23 kDa

  Observed band size: 22 kDa, 37 kDa, 50 kDa

• 

  Immunoprecipitation - Anti-CNTF antibody [EPR24553-104] (ab270992)

  CNTF was immunoprecipitated from 0.35 mg Rat sciatic nerve lysate with ab270992 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab270992 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
  

  Lane 1: Rat sciatic nerve lysate 10ug

  Lane 2: ab270992 IP in Rat sciatic nerve lysate

  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab270992 in rat sciatic nerve lysate

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

  Exposure time: 10 seconds.

  Bands at 40, 75KD are likely to be multimer forms of CNTF (PMID: 9236223).

  All lanes: Immunoprecipitation - Anti-CNTF antibody [EPR24553-104] (ab270992)

  Predicted band size: 23 kDa

  Observed band size: 22 kDa, 37 kDa, 50 kDa

• 

  Immunohistochemistry (Frozen sections) - Anti-CNTF antibody [EPR24553-104] (ab270992)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat sciatic nerve tissue labeling CNTF with ab270992 at 1/100 dilution (4.81 ug/ml) followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on rat sciatic nerve is observed. The nuclear counterstain was DAPI (Blue).
  

  Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• 

  Immunohistochemistry (Frozen sections) - Anti-CNTF antibody [EPR24553-104] (ab270992)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat stomach tissue labeling CNTF with ab270992 at 1/100 dilution (4.81 ug/ml) followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Negative control: No staining on rat stomach (PMID: 12855189) is observed. The nuclear counterstain was DAPI (Blue).
  

  Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• 

  Flow Cytometry (Intracellular) - Anti-CNTF antibody [EPR24553-104] (ab270992)

  Flow cytometric analysis of 4% paraformaldehyde fixed, 0.1% Tween-20 permeabilised Rat primary neural/glia cells labeling CNTF with ab270992 at 1/500 dilution (0.1μg)/ (Right image) compared with an isotype control Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730)/ (Left image) and an unlabeled 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) ab150077), at 1/2000 dilution was used as the secondary antibody.