Overview

  • Product name
    Anti-DDIT3 antibody [9C8]
    See all DDIT3 primary antibodies
  • Description
    Mouse monoclonal [9C8] to DDIT3
  • Host species
    Mouse
  • Tested applications
    Suitable for: WB, IP, ICC, ICC/IF, IHC-Fr, Flow Cyt, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

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

  • Epitope
    ab11419 has been shown to recognize an epitope in the N-terminal region of DDIT3.
  • Positive control
    • ICC/IF: Rat supraoptic nucleus; HeLa cells. WB: HeLa cells treated with tunicamycin (5µg/ml) for 11 hours; Mouse hepatocyte whole cell lysate; Primary human fibroblasts treated with tunicamycin for 10 hours; Mouse 3T3 cells treated with tunicamycin for 24 hours. Flow: HeLa cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab11419 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
WB Use a concentration of 1 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 Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
IHC-Fr 1/100.
Flow Cyt Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

IHC-P Use at an assay dependent concentration. PubMed: 19352619

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

  • Immunostaining of DDIT3 (green) and AVP (red) in the supraoptic nucleus of 3 days DH and 7 days SL rats shows expression of CHOP in the nuclei of AVP MCNs (magnocellular neurons). DDIT3 was stained using ab11419 at 1/200 dilution.

    The rat  brains were removed and post-fixed overnight in 4% (w/v) PFA followed by 30% (w/v) sucrose prepared in PBS to cryoprotect the tissue prior to freezing over liquid nitrogen. Coronal sections (40 μm) of the forebrain were cut on a cryostat and washed in PBS three times. Sections then were blocked in 5% fetal bovine serum prepared in 0.25% (v/v) TritonX/PBS (PBST) for 30 min and then incubated with appropriate primary antibodies at 4°C for 48 hours.

    DH = Complete fluid deprivation.
    SL = Salt loading by drimking 2% salt solution

  • Western blot analysis of HeLa cell lysates using DDIT3 monoclonal antibody (ab11419). HeLa cells were left untreated (-) or treated with tunicamycin (5μg/ml) for 11 hours (+). Membranes containing whole cell extracts were blocked with 5% milk in TBST (blocking buffer) and probed overnight at 4oC with ab11419 (diluted 1:2000 in blocking buffer) followed by a goat anti-mouse IgG-HRP secondary antibody (diluted in blocking buffer) and a chemiluminescent substrate. Membranes were also probed for actin as a loading control.

  • ab11419 staining GADD153 in human HeLa cells by Immunocytochemistry/ Immunofluorescence. The cells were fixed in paraformaldehyde, permeabilised in 0.5% Triton X and then blocked using 3% BSA for 1 hour at 24°C. Samples were then incubated with primary antibody at a 1/100 dilution for 1 hour at 24°C. The secondary antibody used was a goat anti-mouse IgG conjugated to Alexa Fluor® 488 (green) used at a 1/1000 dilution.
    HeLa cells treated with 20uM Tunicamycin for 15 minutes before fixation.

    See Abreview

  • Overlay histogram showing HeLa cells stained with ab11419 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab11419, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b  (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • All lanes : Anti-DDIT3 antibody [9C8] (ab11419) at 1/1000 dilution

    All lanes : Mouse hepatocyte whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-mouse IgG polyclonal at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 19 kDa
    Observed band size: 27 kDa
    why is the actual band size different from the predicted?


    Exposure time: 5 minutes


    Treated with 20µg/ml poly(I:C).

    See Abreview

  • Western blot of endogenous DDIT3 from primary Human fibroblasts using ab11419.
    Lane 1: Untreated cells
    Lane 2: Cells treated with tunicamycin for 10 hours

  • All lanes : Anti-DDIT3 antibody [9C8] (ab11419) at 1/500 dilution (in TBST + 2.5% milk for 16 hours at 4°C)

    Lane 1 : Whole cell lystate of Mouse 3T3 cells
    Lane 2 : Whole cell lystate of Mouse 3T3 cells treated with tunicamycin for 24 hours

    Lysates/proteins at 50 µg per lane.

    Secondary
    All lanes : An HRP-conjugated Goat anti-mouse IgG monoclonal at 1/2000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 19 kDa
    Observed band size: 31 kDa why is the actual band size different from the predicted?


    Exposure time: 2 minutes


    Blocking Step: 5% Milk for 2 hours at 22°C

    See Abreview

  • 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.

References

This product has been referenced in:
  • Leipnitz G  et al. Evaluation of mitochondrial bioenergetics, dynamics, endoplasmic reticulum-mitochondria crosstalk, and reactive oxygen species in fibroblasts from patients with complex I deficiency. Sci Rep 8:1165 (2018). WB ; Human . Read more (PubMed: 29348607) »
  • Xie Q  et al. TAT-fused IP3R-derived peptide enhances cisplatin sensitivity of ovarian cancer cells by increasing ER Ca2+ release. Int J Mol Med 41:809-817 (2018). Read more (PubMed: 29207009) »
See all 83 Publications for this product

Customer reviews and Q&As

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1-10 of 22 Abreviews

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate pH6
Permeabilization
No
Specification
Liver
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

Submitted Oct 22 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate buffer pH6
Permeabilization
No
Specification
brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% formalin

Abcam user community

Verified customer

Submitted Oct 22 2018

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate buffer pH6
Permeabilization
No
Specification
lung
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

Submitted Oct 19 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate buffer pH6
Permeabilization
No
Specification
liver
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% formalin

Abcam user community

Verified customer

Submitted Oct 19 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (brain tissue)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate ph 6
Permeabilization
No
Specification
brain tissue
Blocking step
Normal Goat Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 26°C
Fixative
Formaldehyde

Herr Dr. Markus Kipp

Verified customer

Submitted Aug 17 2018

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Sheep Tissue sections (Intestinal slides, Ileum)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer pH 6.0
Permeabilization
No
Specification
Intestinal slides, Ileum
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 12 2017

Application
ChIP
Sample
Human Cell lysate - whole cell (Heptocyte)
Negative control
Untreated cells
Specification
Heptocyte
Detection step
Semiquantitative PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% formaldehyde
Positive control
Efavirenz

Abcam user community

Verified customer

Submitted Oct 07 2016

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Cell lysate - whole cell (HEPATOCYTES)
Specification
HEPATOCYTES
Treatment
20 ug/ml poly(I:C)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Jul 09 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (SH-SY5Y cell)
Loading amount
100 µg
Specification
SH-SY5Y cell
Treatment
25um cadmium
Gel Running Conditions
Non-reduced Denaturing
Blocking step
Milk as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 08 2013

Application
Western blot
Sample
Human Cell lysate - whole cell (Colon)
Loading amount
250000 cells
Specification
Colon
Treatment
0,5µM Thapsigargin 24h
Gel Running Conditions
Reduced Denaturing (14)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Jul 20 2011

1-10 of 22 Abreviews

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