Anti-DDIT3 antibody [9C8] (ab11419)
Mouse monoclonal DDIT3 antibody [9C8]. Validated in WB, IP, IHC, ICC/IF and tested in Mouse, Rat, Human. Cited in 86 publication(s). Independently reviewed in 22 review(s).
- Datasheet
- References (118)
- Protocols
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/IF, IHC-Frmore 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
- WB: HeLa cells treated with 2ug/ml tunicamycin for 4 hours. 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.
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
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
-
Related Products
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 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. |
|
| IP | Use at an assay dependent concentration. | |
| ICC/IF | Use a concentration of 5 µg/ml. | |
| IHC-Fr | 1/100. |
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
- 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
see all -
Alternative names
- C/EBP homologous protein antibody
- C/EBP Homology Protein antibody
- C/EBP zeta antibody
see all
Images
-
![Western blot - Anti-DDIT3 antibody [9C8] (ab11419)](https://www.abcam.com/ps/products/11/ab11419/Images/ab11419-387590-anti-ddit3-antibody-9c8-western-blot-hela-wc-control-hela-cells-treated-with-2ugml-tunicamycin-for-4-hours-whole-cell-lysate-hepg2-nih3t3.jpg)
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 kDaLanes 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.
-
![Immunocytochemistry/ Immunofluorescence - Anti-DDIT3 antibody [9C8] (ab11419)](https://www.abcam.com/ps/products/11/ab11419/Images/ab11419-319555-anti-ddit3-antibody-9c8-immunofluorescence.jpg)
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)](https://www.abcam.com/ps/products/11/ab11419/Images/ab11419-229338-anti-ddit3-antibody-9c8-western-blot.jpg)
Western blot - Anti-DDIT3 antibody [9C8] (ab11419)This image is courtesy of an anonymous AbreviewAll 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 minutesTreated with 20µg/ml poly(I:C).
-
![Western blot - Anti-DDIT3 antibody [9C8] (ab11419)](https://www.abcam.com/ps/products/11/ab11419/Images/ab11419-18936-anti-ddit3-antibody-9c8-western-blot.jpg)
Western blot - Anti-DDIT3 antibody [9C8] (ab11419)This image is courtesy of an anonymous AbreviewAll 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 -
![Immunocytochemistry/ Immunofluorescence - Anti-DDIT3 antibody [9C8] (ab11419)](https://www.abcam.com/ps/products/11/ab11419/Images/ab11419-203480-anti-ddit3-antibody-9c8-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-DDIT3 antibody [9C8] (ab11419)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.
Protocols
References
This product has been referenced in:
- 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). Read more (PubMed: 31232476) »
- Wang R et al. The Cardiotoxicity Induced by Arsenic Trioxide is Alleviated by Salvianolic Acid A via Maintaining Calcium Homeostasis and Inhibiting Endoplasmic Reticulum Stress. Molecules 24:N/A (2019). Read more (PubMed: 30717322) »
