重组Anti-MEK3抗体[EPR17345-104] (ab195037)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17345-104] to MEK3
- Suitable for: IHC-P, WB, ICC/IF, IP, 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-MEK3抗体[EPR17345-104]
参阅全部 MEK3 一抗 -
描述
兔单克隆抗体[EPR17345-104] to MEK3 -
宿主
Rabbit -
经测试应用
适用于: IHC-P, WB, ICC/IF, IP, Flow Cyt (Intra)more details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HepG2, HeLa and MCF7 whole cell lysates; Human skeletal muscle, mouse spleen and rat spleen lysates. IHC-P: Human skeletal muscle, Human lung adenocarcinoma, mouse skeletal muscle and rat skeletal muscle tissues. ICC/IF: NIH/3T3 cells. IP: HeLa whole cell extract.
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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 (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR17345-104 -
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
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab195037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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IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
1/5000. Detects a band of approximately 39, 34 kDa (predicted molecular weight: 39 kDa).
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ICC/IF |
1/250.
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IP |
1/90.
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Flow Cyt (Intra) |
1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
说明 |
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IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/5000. Detects a band of approximately 39, 34 kDa (predicted molecular weight: 39 kDa). |
ICC/IF
1/250. |
IP
1/90. |
Flow Cyt (Intra)
1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
靶标
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功能
Dual specificity kinase. Is activated by cytokines and environmental stress in vivo. Catalyzes the concomitant phosphorylation of a threonine and a tyrosine residue in the MAP kinase p38. -
组织特异性
Abundant expression is seen in the skeletal muscle. It is also widely expressed in other tissues. -
疾病相关
Note=Defects in MAP2K3 may be involved in colon cancer. -
序列相似性
Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. MAP kinase kinase subfamily.
Contains 1 protein kinase domain. -
翻译后修饰
Autophosphorylated.
Phosphorylation on Ser-218 and Thr-222 by MAP kinase kinase kinases regulates positively the kinase activity.
Yersinia yopJ may acetylate Ser/Thr residues, preventing phosphorylation and activation, thus blocking the MAPK signaling pathway. - Information by UniProt
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数据库链接
- Entrez Gene: 5606 Human
- Entrez Gene: 26397 Mouse
- Entrez Gene: 303200 Rat
- Omim: 602315 Human
- SwissProt: P46734 Human
- SwissProt: O09110 Mouse
- Unigene: 514012 Human
- Unigene: 18494 Mouse
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别名
- AW212142 antibody
- dual specificity mitogen activated protein kinase kinase 3 antibody
- Dual specificity mitogen-activated protein kinase kinase 3 antibody
see all
图片
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: MEK3 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: Jurkat cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab195037 observed at 40 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab195037 was shown to specifically react with MEK3 when MEK3 knockout samples were used. Wild-type and MEK3 knockout samples were subjected to SDS-PAGE. ab195037 and ab8245 (loading control to GAPDH) were both diluted to 1/5000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MEK3 antibody [EPR17345-104] (ab195037)
Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling MEK3 with ab195037 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary at 1/500 dilution. Cytoplasmic and nucleus staining on Human skeletal muscle is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
1:Ben-Levy R,et.al.(1998) Nuclear export of the stress-activated protein kinase p38 mediated by its substrate MAPKAP kinase-2. Curr Biol, 8(19):1049-1057.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embyro fibroblast cells) cells labeling MEK3 with ab195037 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing both nuclear and cytoplasmic staining on NIH/3T3 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab195037 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution. -
Anti-MEK3 antibody [EPR17345-104] (ab195037) at 1/50000 dilution + HepG2 (Human liver hepatocellular carcinoma) whole cell lysates at 10 µg
Secondary
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 39 kDa
Observed band size: 34,39 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Based on the sequence analysis, ab195037 recognizes 3 isoforms of MEK3 ranging from 36kDa to 40KDa. The multi-bands should be MEK3 isoforms.
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Intracellular Flow Cytometry analysis of NIH/3T3 (mouse embryo) labelling MEK3 with purified ab195037 at 1/1000 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Alexa Fluor® 488 goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
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All lanes : Anti-MEK3 antibody [EPR17345-104] (ab195037) at 1/10000 dilution
Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
Lane 2 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 39 kDa
Observed band size: 34,39 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Based on the sequence analysis, ab195037 recognizes 3 isoforms of MEK3 ranging from 36kDa to 40KDa. The multi-bands should be MEK3 isoforms.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MEK3 antibody [EPR17345-104] (ab195037)
Immunohistochemical analysis of paraffin-embedded Human lung adenocarcinoma tissue labeling MEK3 with ab195037 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary at 1/500 dilution. Cytoplasmic and nuclear staining on cancer cells of Human lung adenocarcinoma is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Anti-MEK3 antibody [EPR17345-104] (ab195037) at 1/5000 dilution + Human skeletal muscle lysates at 10 µg
Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 39 kDa
Observed band size: 34,39 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Based on the sequence analysis, ab195037 recognizes 3 isoforms of MEK3 ranging from 36kDa to 40KDa. The multi-bands should be MEK3 isoforms.
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All lanes : Anti-MEK3 antibody [EPR17345-104] (ab195037) at 1/10000 dilution
Lane 1 : Mouse spleen lysates
Lane 2 : Rat spleen lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 39 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
Based on the sequence analysis, ab195037 recognizes 3 isoforms of MEK3 ranging from 36kDa to 40KDa. The multi-bands should be MEK3 isoforms.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MEK3 antibody [EPR17345-104] (ab195037)
Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling MEK3 with ab195037 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary at 1/500 dilution. Cytoplasmic and nuclear staining on mouse skeletal muscle is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MEK3 antibody [EPR17345-104] (ab195037)
Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling MEK3 with ab195037 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary at 1/500 dilution. Cytoplasmic and nuclear staining on rat skeletal muscle is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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MEK3 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell extract with ab195037 at 1/90 dilution. Western blot was performed from the immunoprecipitate using ab195037 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Lane 1: HeLa whole cell extract. Lane 2: PBS instead of HeLa whole cell extract.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Based on the sequence analysis, ab195037 recognizes 3 isoforms of MEK3 ranging from 36kDa to 40KDa. The multi-bands should be MEK3 isoforms.
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 (9)
ab195037 被引用在 9 文献中.
- Deng W et al. RNF150 suppresses papillary thyroid carcinoma with ASK1 ubiquitination presenting a direct target via inactivating p38 signaling axis. Cell Biol Int 47:1198-1208 (2023). PubMed: 36950779
- Huang J et al. Effect of curcumin on regulatory B cells in chronic colitis mice involving TLR/MyD88 signaling pathway. Phytother Res 37:731-742 (2023). PubMed: 36196887
- Xing G et al. MAP2K6 remodels chromatin and facilitates reprogramming by activating Gatad2b-phosphorylation dependent heterochromatin loosening. Cell Death Differ 29:1042-1054 (2022). PubMed: 34815549
- Gartz M et al. Cardiomyocyte-produced miR-339-5p mediates pathology in Duchenne muscular dystrophy cardiomyopathy. Hum Mol Genet 30:2347-2361 (2021). PubMed: 34270708
- Yang X et al. Discovery of the first chemical tools to regulate MKK3-mediated MYC activation in cancer. Bioorg Med Chem 45:116324 (2021). PubMed: 34333394
- Sabry M et al. Tumor- and cytokine-primed human natural killer cells exhibit distinct phenotypic and transcriptional signatures. PLoS One 14:e0218674 (2019). PubMed: 31242243
- Luo S et al. SPAG9/MKK3/p38 axis is a novel therapeutic target for liver cancer. Oncol Rep 41:2329-2336 (2019). PubMed: 30720101
- Yu D et al. Downregulated microRNA-488 enhances odontoblast differentiation of human dental pulp stem cells via activation of the p38 MAPK signaling pathway. J Cell Physiol 234:1442-1451 (2019). PubMed: 30132853
- Sikorski K et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371