Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504]

9 product images

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  Western blot - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] (ab200829)

  Blocking/Dilution buffer: 5% NFDM/TBST.
  

  Based on the sequence analysis, ab200829 recognizes six isoforms within human with predicted Mw's of 56KDa, 55KDa, 48KDa, 45KDa, 48KDa and 49KDa, respectively.

  All lanes: Western blot - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] (ab200829) at 1/10000 dilution

  Lane 1: HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 20 µg

  Lane 2: MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

  Predicted band size: 47 kDa, 56 kDa

  Observed band size: 45-65 kDa

  Exposure time: 30s

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  Immunoprecipitation - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] (ab200829)

  NFIB/NF1B2 + NFIC was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab200829 at 1/70 dilution. Western blot was performed from the immunoprecipitate using ab200829 at 1/5000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.
  

  Lane 1: HeLa whole cell lysate, 10 μg (Input).

  Lane 2: ab200829 IP in HeLa whole cell lysate.

  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab200829 in HeLa whole cell lysate.
  Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  Exposure time: 10 seconds.

  All lanes: Immunoprecipitation - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] (ab200829)

  Observed band size: 45-65 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] (ab200829)

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling NFIB / NF1B2 + NFIC with ab200829 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/500 dilution (green). Nuclear staining on HeLa cell line is observed. The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
  The negative controls are as follows:-
  -ve control 1: ab200829 at 1/1000 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
  -ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] (ab200829)

  Immunohistochemical analysis of paraffin-embedded human cervix carcinoma tissue labeling NFIB / NF1B2 + NFIC with ab200829 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Nuclear staining on Human cervix carcinoma tissue is observed. Counter stained with Hematoxylin.
  

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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  Flow Cytometry (Intracellular) - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] (ab200829)

  Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma)cells labeling NFIB / NF1B2 + NFICwith ab200829 at 1/200 dilution (red) compared with a rabbit monoclonal IgG isotype control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.
  

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] (ab200829)

  Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling NFIB / NF1B2 + NFIC with ab200829 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Nuclear staining on Human breast carcinoma tissue is observed. Counter stained with Hematoxylin.
  

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] (ab200829)

  Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling NFIB / NF1B2 + NFIC with ab200829 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Nuclear staining on mouse cardiac muscle tissue is observed. Counter stained with Hematoxylin.
  

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• 

  Western blot - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] (ab200829)

  Blocking/Dilution buffer:5% NFDM /TBST
  

  Based on the sequence analysis, ab200829 recognizes six isoforms within human with predicted Mw's of 56KDa, 55KDa, 48KDa, 45KDa, 48KDa and 49KDa, respectively.

  All lanes: Western blot - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] (ab200829) at 1/10000 dilution

  All lanes: HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate at 20 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

  Predicted band size: 47 kDa, 56 kDa

  Observed band size: 45-65 kDa

  Exposure time: 3min

• 

  Western blot - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] (ab200829)

  Blocking/Dilution buffer: 5% NFDM/TBST.
  

  Based on the sequence analysis, ab200829 recognizes six isoforms within human with predicted Mw's of 56KDa, 55KDa, 48KDa, 45KDa, 48KDa and 49KDa, respectively.

  All lanes: Western blot - Anti-NFIB / NF1B2 + NFIC/CTF antibody [EPR14504] (ab200829) at 1/2000 dilution

  Lane 1: Mouse heart lysate at 10 µg

  Lane 2: Mouse kidney lysate at 10 µg

  Lane 3: Mouse spleen lysate at 10 µg

  Lane 4: Rat brain lysate at 10 µg

  Lane 5: Rat heart lysate at 10 µg

  Lane 6: Rat spleen lysate at 10 µg

  Lane 7: C6 (Rat glial tumor cells) whole cell lysate at 10 µg

  Lane 8: RAW 264.7(Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

  Lane 9: PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 10 µg

  Lane 10: NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate at 10 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

  Predicted band size: 47 kDa, 56 kDa

  Observed band size: 45-65 kDa

  Exposure time: 1min