Anti-CNTFR antibody [EPR24672-25]

6 product images

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  Western blot - Anti-CNTFR antibody [EPR24672-25] (ab305245)

  CNTFR Western blot staining of SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate using rabbit Anti-CNTFR antibody

  Blocking and diluting buffer and concentration: 5% NFDM /TBST.
  Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used as a GAPDH loading control at 1/1000000 dilution.
  Exposure time:
  Lane 1: 180 seconds
  Lane 2: 20 seconds
  The sensitivity of ab305245 is lower than that of Anti-CNTFR antibody [EPR24672-166] ab283682, we recommend customers to choose Anti-CNTFR antibody [EPR24672-166] ab283682 when performing WB.

  Lane 1: Western blot - Anti-CNTFR antibody [EPR24672-25] (ab305245) at 1/1000 dilution

  Lane 2: Western blot - Anti-CNTFR antibody [EPR24672-166] (Anti-CNTFR antibody [EPR24672-166] ab283682) at 1/1000 dilution

  All lanes: SH-SY5Y (Human neuroblastoma epithelial cell) whole cell 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 1/20000 dilution

  Observed band size: 60 kDa

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  Immunoprecipitation - Anti-CNTFR antibody [EPR24672-25] (ab305245)

  CNTFR was immunoprecipitated from 0.35 mg SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate 10 μg
  with ab305245 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab305245. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
  

  Lane 1: SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate 10 μg

  Lane 2: ab305245 at 1/30 IP in SH-SY5Y whole cell lysate 10 μg

  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab305245 in SH-SY5Y whole cell lysate

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

  This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

  All lanes: Immunoprecipitation - Anti-CNTFR antibody [EPR24672-25] (ab305245) at 1/30 dilution

  All lanes: SH-SY5Y (human neuroblastoma epithelial cell)

  Secondary

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

  Developed using the ECL technique.

  Observed band size: 60 kDa

  Exposure time: 10s

• 

  Western blot - Anti-CNTFR antibody [EPR24672-25] (ab305245)

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

  Negative control: U-87 MG (PMID:23061382)

  This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

  All lanes: Western blot - Anti-CNTFR antibody [EPR24672-25] (ab305245) at 1/1000 dilution

  Lane 1: SH-SY5Y (human neuroblastoma epithelial cell), whole cell lysate at 20 µg

  Lane 2: U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell 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

  Developed using the ECL technique.

  Observed band size: 60 kDa

  Exposure time: 180s

• 

  Western blot - Anti-CNTFR antibody [EPR24672-25] (ab305245)

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

  Low expression: heart, kidney, spleen (PMID:1648265)

  This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

  All lanes: Western blot - Anti-CNTFR antibody [EPR24672-25] (ab305245) at 1/1000 dilution

  Lane 1: Human cerebellum tissue lysate at 20 µg

  Lane 2: Human heart tissue lysate at 20 µg

  Lane 3: Human kidney tissue lysate at 20 µg

  Lane 4: Human spleen tissue 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

  Developed using the ECL technique.

  Observed band size: 60 kDa

  Exposure time: 37s

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  Immunocytochemistry/ Immunofluorescence - Anti-CNTFR antibody [EPR24672-25] (ab305245)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y (human neuroblastoma epithelial cell) cells labelling CNTFR with ab305245 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing mainly membranous staining in SH-SY5Y cell line. Negative control: U-87 MG (PMID: 23061382). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
  
  Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red).
  The Nuclear counterstain was DAPI (Blue).
  Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

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  Flow Cytometry - Anti-CNTFR antibody [EPR24672-25] (ab305245)

  Flow cytometric analysis of U-87 MG (human glioblastoma-astrocytoma epithelial cell, Left) / SH-SY5Y (human neuroblastoma epithelial cell, Right) cells labelling CNTFR with ab305245 at 1/500 dilution (0.1ug) (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. Gated on viable cells.Negative control: U-87 MG (PMID: 23061382).