概述

  • 产品名称Anti-DDIT3抗体[9C8]
    参阅全部 DDIT3 一抗
  • 描述
    小鼠单克隆抗体[9C8] to DDIT3
  • 经测试应用适用于: WB, IP, ICC, ICC/IF, IHC-Fr, Flow Cyt, IHC-Pmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

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

  • 表位ab11419 has been shown to recognize an epitope in the N-terminal region of DDIT3.
  • 阳性对照
    • Primary human fibroblasts treated with tunicamycin for 10 hours.

性能

应用

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.

应用 Ab评论 说明
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

靶标

  • 功能Inhibits the DNA-binding activity of C/EBP and LAP by forming heterodimers that cannot bind DNA.
  • 疾病相关Note=A chromosomal aberration involving DDIT3 is found in a patient with malignant myxoid liposarcoma. Translocation t(12;16)(q13;p11) with FUS.
  • 序列相似性Belongs to the bZIP family.
    Contains 1 bZIP domain.
  • 细胞定位Nucleus.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

Anti-DDIT3 antibody [9C8] 图像



  • Predicted band size : 19 kDa

    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

    Lane 1 : Mouse hepatocyte whole cell lysate
    Lane 2 : Mouse hepatocyte whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    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

    This image is courtesy of an anonymous Abreview

    Treated with 20µg/ml poly(I:C).

    See Abreview



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

    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
    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

    This image is courtesy of an anonymous Abreview

    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.

Anti-DDIT3 antibody [9C8] (ab11419)参考文献

This product has been referenced in:
  • Greenwood M  et al. Transcription Factor CREB3L1 Regulates Endoplasmic Reticulum Stress Response Genes in the Osmotically Challenged Rat Hypothalamus. PLoS One 10:e0124956 (2015). Rat . Read more (PubMed: 25915053) »
  • Varadarajan S  et al. The transrepression arm of glucocorticoid receptor signaling is protective in mutant huntingtin-mediated neurodegeneration. Cell Death Differ 22:1388-96 (2015). Read more (PubMed: 25656655) »

See all 28 Publications for this product

Product Wall

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
Username

Abcam user community

Verified customer

提交于 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
Username

Abcam user community

Verified customer

提交于 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
Username

Abcam user community

Verified customer

提交于 Feb 08 2013

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 purchasin...

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Thank you for contacting Abcam.
I am sorry about the issues that you have been having with ab11419 in IHC-P on human samples. The antibody is covered under our Abpromise for six months and is guaranteed to work in IHC-P on human samples. Based on ...

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Thank you for providing that information.

As I mentioned previously, with the exception of ab79483, we do guarantee that these antibodies should work in western blot on rat tissue. If we cannot resolve the issue you are having with the antibod...

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Thank you for contacting Abcam.

I am sorry that you are having some issues with our antibodies. All of the antibodies you mentioned should work in western blot in various species, however only ab30315 is known to work in IHC-P. Below is a list...

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Thank you for your reply.


I am inclined to believe that the issue is with the samples themselves. It is very unusual for 3 different antibodies to detect bands mainly above 100kDa when the expected MW should be around 25kDa. How long ar...

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Thank you for your reply.

I was wondering if you would clarify a few additional points for me.


How were the samples prepared? How much is loaded on the gel?


What is the molecular weight of each band in the ladd...

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Thank you for contacting Abcam.


I am sorry that you have been experiencing difficulties with these antibodies in WB.


In order to assist you further, I am hoping you can provide some additional details regarding your expe...

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