Recombinant
RabMAb

Recombinant Anti-Fos B antibody [EPR15905] (ab184938)

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Overview

  • Product name

    Anti-Fos B antibody [EPR15905]
    See all Fos B primary antibodies

  • Description

    Rabbit monoclonal [EPR15905] to Fos B

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IP, ICC/IF, WB, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human Fos B aa 1-150. The exact sequence is proprietary.
    Database link: P53539

  • Positive control

    • WB: HeLa whole cell lysate serum starved overnight, then treated with 200nM 12-o-tetradecanoyl phorbol 13-acetate (TPA) for 4hr; NIH/3T3 and C6 whole cell lysate treated with serum, starved overnight, then serum stimulated for 4hr. IHC-P: Mouse hippocampus, mouse kidney, rat hippocampus and rat kidney tissues. ICC/IF: HeLa cells. IP: HeLa whole cell lysate serum starved overnight, then treated with 200nM 12-o-tetradecanoyl phorbol 13-acetate (TPA) for 4hr.

  • 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 ab184938 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
IP 1/50.
ICC/IF 1/500.
WB 1/10000. Detects a band of approximately 38, 48 kDa (predicted molecular weight: 36 kDa).
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

Images

  • Western blot - Anti-Fos B antibody [EPR15905] (ab184938)
    Western blot - Anti-Fos B antibody [EPR15905] (ab184938)
    All lanes : Anti-Fos B antibody [EPR15905] (ab184938) at 1/10000 dilution

    Lane 1 : Untreated HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HeLa whole cell lysate serum starved overnight, then treated with 200nM 12-o-tetradecanoyl phorbol 13-acetate (TPA) for 4hr.

    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: 36 kDa
    Observed band size: 38,48 kDa
    why is the actual band size different from the predicted?


    Exposure time: 5 seconds


    FosB protein exists as two isoforms: full-length FosB (48KD) and a truncated form, FosB2 (38KD, Delta FosB) which lacks the carboxyl-terminal 101 amino acids.

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-Fos B antibody [EPR15905] (ab184938)
    Immunocytochemistry/ Immunofluorescence - Anti-Fos B antibody [EPR15905] (ab184938)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells, untreated or treated with 12-o-tetradecanoyl phorbol 13-acetate (TPA) (200nM) for 4h, labeling Fos B with ab184938 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear staining on HeLa cells. The expression was increaed after treatment with 12-o-tetradecanoyl phorbol 13-acetate (TPA) (200nM) for 4h.

    The nuclear counter stain 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: ab184938 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.

     

  • Western blot - Anti-Fos B antibody [EPR15905] (ab184938)
    Western blot - Anti-Fos B antibody [EPR15905] (ab184938)
    All lanes : Anti-Fos B antibody [EPR15905] (ab184938) at 1/10000 dilution

    Lane 1 : Untreated NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate
    Lane 2 : NIH/3T3 whole cell lysate treated with serum, starved overnight, then serum stimulated for 4hr

    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: 36 kDa
    Observed band size: 38,48 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    FosB protein exists as two isoforms: full-length FosB (48KD) and a truncated form, FosB2 (38KD, Delta FosB) which lacks the carboxyl-terminal 101 amino acids

  • Western blot - Anti-Fos B antibody [EPR15905] (ab184938)
    Western blot - Anti-Fos B antibody [EPR15905] (ab184938)
    All lanes : Anti-Fos B antibody [EPR15905] (ab184938) at 1/10000 dilution

    Lane 1 : Untreated C6 (Rat glial tumor cells) whole cell lysate
    Lane 2 : C6 whole cell lysate treated with serum, starved overnight, then serum stimulated for 4hr

    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: 36 kDa
    Observed band size: 38,48 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    FosB protein exists as two isoforms: full-length FosB (48KD) and a truncated form, FosB2 (38KD, Delta FosB) which lacks the carboxyl-terminal 101 amino acids

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fos B antibody [EPR15905] (ab184938)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fos B antibody [EPR15905] (ab184938)

    Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue labeling Fos B with ab184938 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on mouse hippocampus 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fos B antibody [EPR15905] (ab184938)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fos B antibody [EPR15905] (ab184938)

    Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Fos B with ab184938 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasm staining on mouse kidney 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fos B antibody [EPR15905] (ab184938)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fos B antibody [EPR15905] (ab184938)

    Immunohistochemical analysis of paraffin-embedded rat hippocampus tissue labeling Fos B with ab184938 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on rat hippocampus 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fos B antibody [EPR15905] (ab184938)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fos B antibody [EPR15905] (ab184938)

    Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling Fos B with ab184938 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasm staining on rat kidney 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.

  • Immunoprecipitation - Anti-Fos B antibody [EPR15905] (ab184938)
    Immunoprecipitation - Anti-Fos B antibody [EPR15905] (ab184938)

    Fos B was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate serum starved overnight, then treated with 200nM 12-o-tetradecanoyl phorbol 13-acetate (TPA) for 4hr using ab184938 at 1/50 dilution.

    Western blot was performed from the immunoprecipitate using ab184938 at 1/1000 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 serum starved overnight, then treated with 200nM 12-o-tetradecanoyl phorbol 13-acetate (TPA) for 4hr, 10 µg (Input).

    Lane 2: ab184938 IP in HeLa whole cell lysate serum starved overnight, then treated with 200nM 12-o-tetradecanoyl phorbol 13-acetate (TPA) for 4hr.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab184938 in HeLa whole cell lysate serum starved overnight, then treated with 200nM 12-o-tetradecanoyl phorbol 13-acetate (TPA) for 4hr.

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

References

This product has been referenced in:

  • Niraula A  et al. Corticosterone Production during Repeated Social Defeat Causes Monocyte Mobilization from the Bone Marrow, Glucocorticoid Resistance, and Neurovascular Adhesion Molecule Expression. J Neurosci 38:2328-2340 (2018). Read more (PubMed: 29382712) »
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Immunohistochemistry (Frozen sections) abreview for Anti-Fos B antibody [EPR15905]

 Excellent
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
Yes - Triton X-100
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde
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Submitted Jul 06 2016

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