製品の概要

  • 製品名
    Anti-DDIT3 antibody [9C8]
    DDIT3 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [9C8] to DDIT3
  • 由来種
    Mouse
  • アプリケーション
    適用あり: WB, IP, ICC/IF, IHC-Fr, IHC-Pmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

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

  • エピトープ
    ab11419 has been shown to recognize an epitope in the N-terminal region of DDIT3.
  • ポジティブ・コントロール
    • WB: HeLa cells treated with 2ug/ml tunicamycin for 4 hours. ICC/IF: HeLa (untreated and tunicamycin-treated). IHC-P: Human normal testis and pancreas adenocarcinoma tissues.
  • 特記事項

    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.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.
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.

ターゲット情報

  • 機能
    Inhibits the DNA-binding activity of C/EBP and LAP by forming heterodimers that cannot bind DNA.
  • 関連疾患
    Note=A chromosomal aberration involving DDIT3 is found in a patient with malignant myxoid liposarcoma. Translocation t(12;16)(q13;p11) with FUS.
  • 配列類似性
    Belongs to the bZIP family.
    Contains 1 bZIP domain.
  • 細胞内局在
    Nucleus.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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

画像

  • Lanes 1-2 : Anti-DDIT3 antibody [9C8] (ab11419) at 5 µg/ml
    Lanes 3-4 : Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) at 1/10000 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.

  • 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 (pH 6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab11419, 5 µg/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 hematoxylin 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

  • 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).

  • 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 (pH 6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab11419, 5 µg/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 hematoxylin 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

  • 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

  • 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

  • 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.

参考文献

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 86 Publications for this product

レビューと Q&A

1-10 of 38 Abreviews or Q&A

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

投稿 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

投稿 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

投稿 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

投稿 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

投稿 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

投稿 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

投稿 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

投稿 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

投稿 Feb 08 2013

Answer

Thank you for your reply.
Your credit note ID is ***.
I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.
Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.
The credit note ID is for your reference only and does not automatically guarantee the credit.
I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

Read More

1-10 of 38 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

登録