Overview

  • Product name

    Anti-DDIT3 antibody [9C8] - BSA and Azide free
    See all DDIT3 primary antibodies
  • Description

    Mouse monoclonal [9C8] to DDIT3 - BSA and Azide free
  • Host species

    Mouse
  • Tested applications

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

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Other Immunogen Type corresponding to DDIT3. A bacterially expressed, mouse DDIT3 fusion protein.

  • Epitope

    ab233121 has been shown to recognize an epitope in the N-terminal region of DDIT3.
  • Positive control

    • WB: HeLa cells treated with 2ug/ml tunicamycin for 4 hours. ICC/IF: HeLa (untreated and tunicamycin-treated); SKNSH cells treated with deltamethrin. IHC-P: Human normal testis and pancreas adenocarcinoma tissues.
  • General notes

    This antibody clone is manufactured by Abcam. If you require a different buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com.

    ab233121 is a PBS only format of ab11419.

Properties

Applications

Our Abpromise guarantee covers the use of ab233121 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
IHC-Fr 1/100.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB Use a concentration of 5 µg/ml. Detects a band of approximately 31 kDa (predicted molecular weight: 19 kDa).

Please note that under normal cellular conditions this protein is not expressed in detectable levels, but is highly upregulated during times of cellular/ER stress. It is strongly recommended to run a positive control along your samples to confirm the expression levels of protein.

IP Use at an assay dependent concentration.
ICC/IF Use a concentration of 5 µg/ml.

Target

  • Function

    Inhibits the DNA-binding activity of C/EBP and LAP by forming heterodimers that cannot bind DNA.
  • Involvement in disease

    Note=A chromosomal aberration involving DDIT3 is found in a patient with malignant myxoid liposarcoma. Translocation t(12;16)(q13;p11) with FUS.
  • Sequence similarities

    Belongs to the bZIP family.
    Contains 1 bZIP domain.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • C/EBP homologous protein antibody
    • C/EBP Homology Protein antibody
    • C/EBP zeta antibody
    • C/EBP-homologous protein 10 antibody
    • C/EBP-homologous protein antibody
    • CCAAT/enhancer binding protein homologous protein antibody
    • CEBPZ antibody
    • CHOP 10 antibody
    • CHOP antibody
    • CHOP-10 antibody
    • CHOP10 antibody
    • DDIT 3 antibody
    • DDIT-3 antibody
    • Ddit3 antibody
    • DDIT3_HUMAN antibody
    • DNA Damage Inducible Transcript 3 antibody
    • DNA damage-inducible transcript 3 protein antibody
    • GADD 153 antibody
    • GADD153 antibody
    • Growth Arrest and DNA Damage Inducible Protein 153 antibody
    • Growth arrest and DNA damage inducible protein GADD153 antibody
    • Growth arrest and DNA damage-inducible protein GADD153 antibody
    • MGC4154 antibody
    see all

Images

  • Lanes 1-2 : Anti-DDIT3 antibody [9C8] (ab11419) at 5 µg/ml
    Lanes 3-4 : Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) at 1/1000 dilution

    Lanes 1 & 3 : HeLa whole cell lysate
    Lanes 2 & 4 : HeLa cells treated with 2ug/ml tunicamycin for 4 hours, whole cell lysate

    Lysates/proteins at 40 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 19 kDa
    Observed band size: 28 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 37 kDa (possible Loading Control)



    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 55 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before ab11419 and ab181602 (Rabbit anti GAPDH) were incubated overnight at 4°C at a 5ug/ml concentration and 1/10000 dilution respectively. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, and sodium azide (ab11419).

  • IHC image of DDIT3 staining in a section of formalin-fixed paraffin-embedded human pancreas adenocarcinoma* performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab11419, 5ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, and sodium azide (ab11419).

  • ab11419 staining DDIT3 in HeLa cells +/- Tunicamycin (1.5μM, 6 hours). 

    The cells were fixed with 4% PFA (10min), permeabilized with 0.1% Triton-X for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab11419 at 5μg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control, at 1/1000 dilution. Cells were then incubated with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (shown in green) and ab150084, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolor red). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, and sodium azide (ab11419).

  • IHC image of DDIT3 staining in a section of formalin-fixed paraffin-embedded normal human testis* performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab11419, 5ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, and sodium azide (ab11419).

  • ab11419 staining DDIT3 in SKNSH cells treated with deltamethrin (ab141019), by ICC/IF. Increase of DDIT3 expression correlates with increased concentration of deltamethrin, as described in literature.
    The cells were incubated at 37°C for 48 hours in media containing different concentrations of ab141019 (deltamethrin) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab11419 (10 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight® 488 anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, and sodium azide (ab11419).

References

ab233121 has not yet been referenced specifically in any publications.

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