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
    • Primary human fibroblasts treated with tunicamycin for 10 hours.

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

  • 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:
  • Wang X  et al. Lyn kinase represses mucus hypersecretion by regulating IL-13-induced endoplasmic reticulum stress in asthma. EBioMedicine 15:137-149 (2017). WB, IHC-P ; Mouse, Human . Read more (PubMed: 28024734) »
  • Wu Z  et al. Silencing of both ATF4 and PERK inhibits cell cycle progression and promotes the apoptosis of differentiating chondrocytes. Int J Mol Med 40:101-111 (2017). WB, IHC ; Mouse . Read more (PubMed: 28498443) »

See all 54 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

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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
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Abcam user community

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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
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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
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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
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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
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Submitted Jul 20 2011

Application
Western blot
Sample
Cow Cell lysate - whole cell (vascular endothelial cells)
Loading amount
20 µg
Specification
vascular endothelial cells
Gel Running Conditions
Reduced Denaturing (15% gel)
Blocking step
Milk as blocking agent for 20 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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Dr. Yael Riahi

Verified customer

Submitted Feb 03 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cultured Cells (MEF)
Specification
MEF
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton-X
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
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Submitted Oct 12 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cultured Cells (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton-X
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
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Verified customer

Submitted Oct 12 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (THP-1)
Loading amount
50 µg
Specification
THP-1
Treatment
Tunicamycin for 24hrs
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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Submitted Oct 07 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HELA)
Loading amount
50 µg
Specification
HELA
Treatment
20uM Tunicamycin for 4hrs
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
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Submitted Oct 05 2010

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