重组Anti-DUSP4抗体[EPR19881] (ab216576)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19881] to DUSP4
- Suitable for: ICC/IF, IP, WB, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-DUSP4抗体[EPR19881]
参阅全部 DUSP4 一抗 -
描述
兔单克隆抗体[EPR19881] to DUSP4 -
宿主
Rabbit -
经测试应用
适用于: ICC/IF, IP, WB, Flow Cyt (Intra)more details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: MDA-MB-231, A549, Wild-type A549, SK-BR-3, HCT 116, RAW 264.7, PC-12, MOLT-4 and C6 whole cell lysates; Human breast cancer lysate. ICC/IF: A549 and MDA-MB-231 cells. Flow Cyt (intra): MDA-MB-231 cells, A549 cells. IP: MDA-MB-231 whole cell lysate.
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常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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 -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR19881 -
同种型
IgG -
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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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab216576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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ICC/IF |
Use a concentration of 1 µg/ml.
This product gave a positive signal in A549 (DUSP4 knockout A549 cells used as a negative control) fixed with 100% methanol (5 min). |
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IP |
1/30.
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WB |
1/1000. Detects a band of approximately 43 kDa (predicted molecular weight: 43 kDa).
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Flow Cyt (Intra) |
1/60.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
说明 |
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ICC/IF
Use a concentration of 1 µg/ml. This product gave a positive signal in A549 (DUSP4 knockout A549 cells used as a negative control) fixed with 100% methanol (5 min). |
IP
1/30. |
WB
1/1000. Detects a band of approximately 43 kDa (predicted molecular weight: 43 kDa). |
Flow Cyt (Intra)
1/60. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
靶标
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功能
Regulates mitogenic signal transduction by dephosphorylating both Thr and Tyr residues on MAP kinases ERK1 and ERK2. -
序列相似性
Belongs to the protein-tyrosine phosphatase family. Non-receptor class dual specificity subfamily.
Contains 1 rhodanese domain.
Contains 1 tyrosine-protein phosphatase domain. -
翻译后修饰
Phosphorylation in the C-terminus by ERK1/2 inhibits proteasomal degradation and stabilizes the protein. -
细胞定位
Nucleus. - Information by UniProt
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数据库链接
- Entrez Gene: 1846 Human
- Entrez Gene: 319520 Mouse
- Entrez Gene: 60587 Rat
- Omim: 602747 Human
- SwissProt: Q13115 Human
- SwissProt: Q8BFV3 Mouse
- SwissProt: Q62767 Rat
- Unigene: 417962 Human
see all -
别名
- Dual specificity phosphatase 4 antibody
- Dual specificity protein phosphatase 4 antibody
- Dual specificity protein phosphatase hVH2 antibody
see all
图片
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Flow cytometry overlay histogram showing left wild-type A549 positive cells and right negative DUSP4 knockout A549 stained with ab216576 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab216576) (1x 106 in 100μl at 1.0 μg/ml (1/1990)) for 30min at 22°C.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C
Isotype control antibody Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (black line) was used at the same concentration and conditions as the primary antibody. Unlabelled sample was also used as a control (blue line).
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
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All lanes : Anti-DUSP4 antibody [EPR19881] (ab216576) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : DUSP4 knockout A549 cell lysate
Lane 3 : A549 cell lysate
Lane 4 : MOLT-4 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 43 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab216576 observed at 40 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab216576 was shown to react with DUSP4 in wild-type A549 cells in Western blot with loss of signal observed in DUSP4 knockout cell line ab273859 (DUSP4 knockout cell lysate ab273813). Wild-type A549 and DUSP4 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab216576 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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All lanes : Anti-DUSP4 antibody [EPR19881] (ab216576) at 1/1000 dilution
Lane 1 : MDA-MB-231 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg
Lane 2 : A549 (Human lung carcinoma cell line) whole cell lysate at 10 µg
Lane 3 : SK-BR-3 (Human mammary gland adenocarcinoma cell line) whole cell lysate at 10 µg
Lane 4 : HCT 116 (Human colorectal carcinoma cell line) whole cell lysate at 10 µg
Secondary
Lanes 1-2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lanes 3-4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 43 kDa
Observed band size: 43 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1/2:3 minutes; Lane 3: 30 seconds; Lane 4: 1 second.
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ab216576 staining DUSP4 in wild-type A549 cells, with negative expression in DUSP4 knockout A549 cells. The cells were fixed with 100% methanol (5 min), permeabilised with 0.1% Triton x-100 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 ab216576 at 1 μg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin at 0.5 μg/ml. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150119, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 647), pre-adsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A549 (Human lung carcinoma cell line) cells labeling DUSP4 with ab216576 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing nuclear staining on A549 cell line.
The nuclear counterstain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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Anti-DUSP4 antibody [EPR19881] (ab216576) at 1/1000 dilution + Human breast cancer lysate at 10 µg
Secondary
VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 43 kDa
Observed band size: 43 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-DUSP4 antibody [EPR19881] (ab216576) at 1/1000 dilution
Lane 1 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 3 : C6 (Rat glial tumor cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 43 kDa
Observed band size: 43 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1:3 minutes; Lane 2/3: 15 seconds.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MDA-MB-231 (Human breast adenocarcinoma cell line) cells labeling DUSP4 with ab216576 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing nuclear staining on MDA-MB-231 cell line.
The nuclear counterstain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed MDA-MB-231 (Human breast adenocarcinoma cell line) cells labeling DUSP4 with ab216576 at 1/60 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
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DUSP4 was immunoprecipitated from 0.35mg of MDA-MB-231 (Human breast adenocarcinoma cell line) whole cell lysate with ab216576 at 1/30 dilution.
Western blot was performed from the immunoprecipitate using ab216576 at 1/500 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1,000 dilution
Lane 1: MDA-MB-231 whole cell lysate, 10µg (Input).
Lane 2: ab216576 IP in MDA-MB-231 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab216576 in MDA-MB-231 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (10)
ab216576 被引用在 10 文献中.
- Clemm von Hohenberg K et al. Cyclin B/CDK1 and Cyclin A/CDK2 phosphorylate DENR to promote mitotic protein translation and faithful cell division. Nat Commun 13:668 (2022). PubMed: 35115540
- Tang Y et al. ING4 Promotes Stemness Enrichment of Human Renal Cell Carcinoma Cells Through Inhibiting DUSP4 Expression to Activate the p38 MAPK/type I IFN-Stimulated Gene Signaling Pathway. Front Pharmacol 13:845097 (2022). PubMed: 35496267
- Yang C et al. Silencing circFTO inhibits malignant phenotype through modulating DUSP4 expression in clear cell renal cell carcinoma. Cell Death Discov 8:392 (2022). PubMed: 36127345
- Pan AL et al. Dual-Specificity Protein Phosphatase 4 (DUSP4) Overexpression Improves Learning Behavior Selectively in Female 5xFAD Mice, and Reduces β-Amyloid Load in Males and Females. Cells 11:N/A (2022). PubMed: 36497141
- Cheng G et al. The microRNA-429/DUSP4 axis regulates the sensitivity of colorectal cancer cells to nintedanib. Mol Med Rep 23:N/A (2021). PubMed: 33495832
- Zhao W & Wang Q Knockdown of TRIM9 attenuates irinotecan‑induced intestinal mucositis in IEC‑6 cells by regulating DUSP6 expression via the P38 pathway. Mol Med Rep 24:N/A (2021). PubMed: 34676875
- Yan J et al. Dusp4 Contributes to Anesthesia Neurotoxicity via Mediated Neural Differentiation in Primates. Front Cell Dev Biol 8:786 (2020). PubMed: 32974341
- Denhez B et al. Diabetes-Induced DUSP4 Reduction Promotes Podocyte Dysfunction and Progression of Diabetic Nephropathy. Diabetes 68:1026-1039 (2019). PubMed: 30862678
- Ding Y et al. Novel clinicopathological and molecular characterization of metanephric adenoma: a study of 28 cases. Diagn Pathol 13:54 (2018). PubMed: 30111351
- Edwards NA et al. Knockdown of p66Shc Alters Lineage-Associated Transcription Factor Expression in Mouse Blastocysts. Stem Cells Dev 27:1479-1493 (2018). PubMed: 30091687