Recombinant
RabMAb

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

Overview

  • Product name

    Anti-NFIB / NF1B2 + NFIC / CTF antibody [EPR14504]
    See all NFIB / NF1B2+NFIC primary antibodies
  • Description

    Rabbit monoclonal [EPR14504] to NFIB / NF1B2 + NFIC / CTF
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt, IP, WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human NFIC aa 350-450. The exact sequence is proprietary.
    Database link: P08651

  • Positive control

    • WB: HeLa, HepG2, MCF7, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Mouse heart, kidney and spleen lysates; Rat brain, heart and spleen lysates. IHC-P: Human cervix carcinoma and breast carcinoma, and mouse cardiac muscle tissues. ICC/IF: HeLa cells. Flow Cyt: HeLa cells. IP: HeLa whole cell lysate.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab200829 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt 1/200.
IP 1/70.
WB 1/2000. Detects a band of approximately 45-65 kDa (predicted molecular weight: 56, 47 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/1000.

Target

Images

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

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 2 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 56, 47 kDa
    Observed band size: 45-65 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    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.

  • 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) (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 ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and 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 ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

  • 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) (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) (ab97051) at 1/500 dilution.

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

  • Flow cytometric analysis of 2% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling NFIB / NF1B2 + NFIC with ab200829 at 1/200 dilution (red) compared with a rabbit monoclonal IgG 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.

  • 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 (ab172730) instead of ab200829 in HeLa whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds.

  • 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) (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) (ab97051) at 1/500 dilution.

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

  • 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) (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) (ab97051) at 1/500 dilution.

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

  • Anti-NFIB / NF1B2 + NFIC / CTF antibody [EPR14504] (ab200829) at 1/10000 dilution + HepG2 (Human liver hepatocellular carcinoma cell line) at 20 µg

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

    Predicted band size: 56, 47 kDa
    Observed band size: 45-65 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


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

    Lane 1 : Mouse heart lysate
    Lane 2 : Mouse kidney lysate
    Lane 3 : Mouse spleen lysate
    Lane 4 : Rat brain lysate
    Lane 5 : Rat heart lysate
    Lane 6 : Rat spleen lysate
    Lane 7 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 8 : RAW 264.7(Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 9 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 10 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 56, 47 kDa
    Observed band size: 45-65 kDa why is the actual band size different from the predicted?


    Exposure time: 1 minute


    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.

References

ab200829 has not yet been referenced specifically in any publications.

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