Anti-DDIT3抗体[9C8] - BSA and Azide free (ab233121)
Key features and details
- Mouse monoclonal [9C8] to DDIT3 - BSA and Azide free
- Suitable for: WB, ICC/IF
- Knockout validated
- Reacts with: Mouse, Human
- Isotype: IgG2b
概述
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产品名称
Anti-DDIT3抗体[9C8] - BSA and Azide free
参阅全部 DDIT3 一抗 -
描述
小鼠单克隆抗体[9C8] to DDIT3 - BSA and Azide free -
宿主
Mouse -
经测试应用
适用于: WB, ICC/IFmore details -
种属反应性
与反应: Mouse, Human
预测可用于: Rat -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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表位
ab233121 has been shown to recognize an epitope in the N-terminal region of DDIT3. -
阳性对照
- WB: SW480 cell lysates, HeLa cells treated with 2ug/ml tunicamycin for 4 hours. ICC/IF: HeLa (untreated and tunicamycin-treated).
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常规说明
ab233121 is the carrier-free version of ab11419.
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at +4°C. Do Not Freeze. -
存储溶液
Constituent: PBS -
无载体
是 -
Concentration information loading...
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纯度
Protein G purified -
克隆
单克隆 -
克隆编号
9C8 -
同种型
IgG2b -
轻链类型
kappa -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab233121于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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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. |
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ICC/IF |
Use a concentration of 5 µg/ml.
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说明 |
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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. |
ICC/IF
Use a concentration of 5 µg/ml. |
靶标
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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
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数据库链接
- 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 -
别名
- C/EBP homologous protein antibody
- C/EBP Homology Protein antibody
- C/EBP zeta antibody
see all
图片
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All lanes : Anti-DDIT3 antibody [9C8] (ab11419) at 5 µg/ml
Lane 1 : Wild-type HeLa Vehicle Control Tunicamycin (20ug/mL, 4h) cell lysate
Lane 2 : Wild-type HeLa Treated Tunicamycin (20ug/mL, 4h) cell lysate
Lane 3 : DDIT3 knockout HeLa Vehicle Control Tunicamycin (20ug/mL, 4h) cell lysate
Lane 4 : DDIT3 knockout HeLa Treated Tunicamycin (20ug/mL, 4h) cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 19 kDa
Observed band size: 25 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-DDIT3 antibody [9C8] staining at 5 µg/ml, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab11419 was shown to bind specifically to DDIT3. A band was observed at 25 kDa in wild-type y cell lysates with no signal observed at this size in DDIT3 knockout cell line ab265760 (knockout cell lysate ab256889). To generate this image, wild-type and DDIT3 knockout y cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution
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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?This data was developed using the same antibody clone in a different buffer formulation (ab11419).
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.
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ab11419 staining DDIT3 in HeLa cells +/- Tunicamycin (1.5μM, 6 hours).
The cells were fixed with 4% PFA (10min), permeabilized with 0.1% Triton-X for 5 minutes 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 labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, and sodium azide (ab11419).
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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
Observed band size: 27 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab11419).
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.
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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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, and sodium azide (ab11419).
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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Datasheet download
文献 (3)
ab233121 被引用在 3 文献中.
- Yao Y et al. circ_014260/miR-384/THBS1 aggravates spinal cord injury in rats by promoting neuronal apoptosis and endoplasmic reticulum stress. Am J Transl Res 14:518-533 (2022). PubMed: 35173872
- Wang S et al. Mechanism of Chronic Stress-Induced Glutamatergic Neuronal Damage in the Basolateral Amygdaloid Nucleus. Anal Cell Pathol (Amst) 2021:8388527 (2021). PubMed: 34858775
- Bagchi AK et al. Endoplasmic Reticulum Stress Promotes iNOS/NO and Influences Inflammation in the Development of Doxorubicin-Induced Cardiomyopathy. Antioxidants (Basel) 10:N/A (2021). PubMed: 34943000