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    ddit3-antibody-9c8-ab11419.pdf

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Epigenetics and Nuclear Signaling Transcription Other factors
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Validated using a knockout cell line

Anti-DDIT3 antibody [9C8] (ab11419)

  • Datasheet
  • SDS
Reviews (29)Q&A (16)References (161)

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Western blot - Anti-DDIT3 antibody [9C8] (ab11419)
  • Immunocytochemistry/ Immunofluorescence - Anti-DDIT3 antibody [9C8] (ab11419)
  • Western blot - Anti-DDIT3 antibody [9C8] (ab11419)
  • Western blot - Anti-DDIT3 antibody [9C8] (ab11419)
  • Western blot - Anti-DDIT3 antibody [9C8] (ab11419)
  • Immunocytochemistry/ Immunofluorescence - Anti-DDIT3 antibody [9C8] (ab11419)

Key features and details

  • Mouse monoclonal [9C8] to DDIT3
  • Suitable for: WB, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Human
  • Isotype: IgG2b

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Knockout
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Human DDIT3 knockout SW480 cell line (ab269585)

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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, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • 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

    • WB: SW480 cell lysates, HeLa cells treated with 2ug/ml tunicamycin for 4 hours, NIH3T3 cell lysate. ICC/IF: HeLa (untreated and tunicamycin-treated).
  • General notes

    Western blot protocol advice:

    DDIT3 is upregulated as a result of cellular or ER stress. It is strongly recommended to run a positive control (such as tunicamycin treated cell lysates) alongside your samples to confirm the protein expression level.
    For blocking, we recommend using 3% milk for 1 hour. Please see the WB image legend for more protocol information.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.02% Sodium azide
    Constituents: PBS, 6.97% L-Arginine
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Monoclonal
  • Clone number

    9C8
  • Isotype

    IgG2b
  • Light chain type

    kappa
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Stem Cells
    • Embryonic Stem Cells
    • Intracellular
    • Developmental Biology
    • Embryogenesis
    • Embryonic stem cells
    • Surface molecules
    • Metabolism
    • Types of disease
    • Diabetes
    • Metabolism
    • Types of disease
    • Obesity
    • Metabolism
    • Types of disease
    • Metabolic disorders

Associated products

  • Alternative Versions

    • Alexa Fluor® 647 Anti-DDIT3 antibody [9C8] (ab233045)
    • Anti-DDIT3 antibody [9C8] - BSA and Azide free (ab233121)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Isotype control

    • Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control (ab170192)
  • KO cell lines

    • Human DDIT3 knockout SW480 cell line (ab269585)
  • Related Products

    • Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)
    • Deltamethrin, protein phosphatase 2B inhibitor (ab141019)

Applications

The Abpromise guarantee

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 (13)
Use a concentration of 5 µg/ml. Detects a band of approximately 31 kDa (predicted molecular weight: 19 kDa).

DDIT3 is upregulated as a result of cellular or ER stress. It is strongly recommended to run a positive control (such as tunicamycin treated cell lysates) alongside your samples to confirm the protein expression level.

For blocking, we recommend using 3% milk for 1 hour. Please see the WB image legend for more protocol information.

ICC/IF (4)
Use a concentration of 5 µg/ml.
Notes
WB
Use a concentration of 5 µg/ml. Detects a band of approximately 31 kDa (predicted molecular weight: 19 kDa).

DDIT3 is upregulated as a result of cellular or ER stress. It is strongly recommended to run a positive control (such as tunicamycin treated cell lysates) alongside your samples to confirm the protein expression level.

For blocking, we recommend using 3% milk for 1 hour. Please see the WB image legend for more protocol information.

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.
  • Target information above from: UniProt accession P35638 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 1649 Human
    • Entrez Gene: 13198 Mouse
    • Entrez Gene: 29467 Rat
    • Omim: 126337 Human
    • SwissProt: P35638 Human
    • SwissProt: P35639 Mouse
    • SwissProt: Q62857 Rat
    • Unigene: 505777 Human
    • Unigene: 728989 Human
    • Unigene: 110220 Mouse
    • Unigene: 11183 Rat
    see all
  • 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 - Anti-DDIT3 antibody [9C8] (ab11419)
    Western blot - Anti-DDIT3 antibody [9C8] (ab11419)
    All lanes : Anti-DDIT3 antibody [9C8] (ab11419) at 5 µg/ml

    Lane 1 : Wild-type SW480 cell lysate
    Lane 2 : DDIT3 knockout SW480 cell lysate
    Lane 3 : Untreated HeLa cell lysate
    Lane 4 : HeLa + DMSO control cell lysate
    Lane 5 : HeLa + tunicamycin (20ug/mL,4 hours) cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 19 kDa
    Observed band size: 26 kDa why is the actual band size different from the predicted?



    Lanes 1 - 5: Merged signal (red and green). Green - ab11419 observed at 26 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.

    ab11419 was shown to react with DDIT3 in wild-type SW480 cells in western blot with loss of signal observed in DDIT3 knockout sample. Wild-type and DDIT3 knockout SW480 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab11419 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at 5 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-DDIT3 antibody [9C8] (ab11419)
    Immunocytochemistry/ Immunofluorescence - Anti-DDIT3 antibody [9C8] (ab11419)

    ab11419 staining DDIT3 in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells +/- Tunicamycin (1.5μM, 6 hours). 

    The cells were fixed with 4% PFA (10 min), permeabilized with 0.1% Triton-X for 5 mins 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 labeled with DAPI (shown in blue).

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

  • Western blot - Anti-DDIT3 antibody [9C8] (ab11419)
    Western blot - Anti-DDIT3 antibody [9C8] (ab11419)
    All lanes : Anti-DDIT3 antibody [9C8] (ab11419) at 5 µg/ml

    Lane 1 : HeLa w/c control cell lysate at 40 µg
    Lane 2 : HeLa cells treated with 2ug/ml tunicamycin for 4 hours, whole cell lysate cell lysate at 40 µg
    Lane 3 : HeLa cells treated with 20ug/ml tunicamycin for 4 hours, whole cell lysate cell lysate at 40 µg
    Lane 4 : HepG2 cell lysate at 20 µg
    Lane 5 : NIH3T3 cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 19 kDa



    Lanes 1 - 5: Merged signal (red and green). Green - ab11419 observed at 27 kDa. Red - loading control, Rabbit anti Actin observed at 42kDa.

    ab11419 was shown to react with DDIT3 in western blot. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab11419 and Rabbit anti Actin overnight at 4°C at 5 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-DDIT3 antibody [9C8] (ab11419)
    Western blot - Anti-DDIT3 antibody [9C8] (ab11419)This image is courtesy of an anonymous Abreview
    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 - Anti-DDIT3 antibody [9C8] (ab11419)
    Western blot - Anti-DDIT3 antibody [9C8] (ab11419)This image is courtesy of an anonymous Abreview
    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

  • Immunocytochemistry/ Immunofluorescence - Anti-DDIT3 antibody [9C8] (ab11419)
    Immunocytochemistry/ Immunofluorescence - Anti-DDIT3 antibody [9C8] (ab11419)

    ab11419 staining DDIT3 in SK-N-SH (human neuroblastoma cell line) 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.

Protocols

  • Flow cytometry protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (161)

Publishing research using ab11419? Please let us know so that we can cite the reference in this datasheet.

ab11419 has been referenced in 161 publications.

  • Jiang L  et al. Inhibition of microRNA-103 attenuates inflammation and endoplasmic reticulum stress in atherosclerosis through disrupting the PTEN-mediated MAPK signaling. J Cell Physiol 235:380-393 (2020). PubMed: 31232476
  • Chen Y  et al. Iridoid glycosides from Radix Scrophulariae attenuates focal cerebral ischemia-reperfusion injury via inhibiting endoplasmic reticulum stress-mediated neuronal apoptosis in rats. Mol Med Rep 21:131-140 (2020). PubMed: 31746404
  • Shi Z  et al. Attenuation of intermittent hypoxia-induced apoptosis and fibrosis in pulmonary tissues via suppression of ER stress activation. BMC Pulm Med 20:92 (2020). PubMed: 32299413
  • Tan X  et al. Fibroblast Growth Factor 2 Attenuates Renal Ischemia-Reperfusion Injury via Inhibition of Endoplasmic Reticulum Stress. Front Cell Dev Biol 8:147 (2020). PubMed: 32266254
  • Tian F  et al. KLF6 Induces Apoptosis in Human Lens Epithelial Cells Through the ATF4-ATF3-CHOP Axis. Drug Des Devel Ther 14:1041-1055 (2020). PubMed: 32210535
View all Publications for this product

Customer reviews and Q&As

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1-10 of 29 Abreviews

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-DDIT3 antibody [9C8]

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrat, pH: 6.0
Permeabilization
No
Specification
brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Paraformaldehyde
Read More

Miss. Katerina Manzhula

Verified customer

Submitted Nov 19 2020

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-DDIT3 antibody [9C8]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH=9.0
Permeabilization
No
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Oct 21 2020

Western blot abreview for Anti-DDIT3 antibody [9C8]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Prostate cancer cell line)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
25 µg
Treatment
100 uM Cobalt Chloride
Specification
Prostate cancer cell line
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Dec 27 2019

Immunocytochemistry/ Immunofluorescence abreview for Anti-DDIT3 antibody [9C8]

Average
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Hepatocytes)
Permeabilization
No
Specification
Hepatocytes
Blocking step
Serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 10% · Temperature: 4°C
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

Submitted Jul 12 2019

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-DDIT3 antibody [9C8]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Spleen)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Ph:6.0
Permeabilization
No
Specification
Spleen
Blocking step
BSA as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 10%
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

Submitted Jul 01 2019

Western blot abreview for Anti-DDIT3 antibody [9C8]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Liver)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (15)
Loading amount
30 µg
Treatment
200µM Palmitate
Specification
Liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Jun 27 2019

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-DDIT3 antibody [9C8]

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate PH:6.0
Permeabilization
No
Specification
Liver
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

Submitted Jun 16 2019

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-DDIT3 antibody [9C8]

Good
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Oct 22 2018

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-DDIT3 antibody [9C8]

Average
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Oct 22 2018

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-DDIT3 antibody [9C8]

Good
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Oct 19 2018

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