重组Anti-MEF2D抗体[EPR24993-10] (ab282731)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR24993-10] to MEF2D
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-MEF2D抗体[EPR24993-10]
参阅全部 MEF2D 一抗 -
描述
兔单克隆抗体[EPR24993-10] to MEF2D -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), ICC/IF, WB, IHC-Pmore details
不适用于: IP -
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: THP-1, RAW 264.7, 2.4G2, His-tagged human MEF2D recombinant protein lysates. IHC-P: Mouse testis, Mouse cerebrum, Rat colon, and Rat cerebrum tissues. ICC/IF: THP-1, RAW 264.7 cells. Flow Cyt(Intra): THP-1, RAW 264.7 cells.
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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. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59.94% 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
EPR24993-10 -
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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Isotype control
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab282731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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Flow Cyt (Intra) |
1/500.
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ICC/IF |
1/50.
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WB |
1/1000. Predicted molecular weight: 56 kDa.
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IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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说明 |
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Flow Cyt (Intra)
1/500. |
ICC/IF
1/50. |
WB
1/1000. Predicted molecular weight: 56 kDa. |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
靶标
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功能
Transcriptional activator which binds specifically to the MEF2 element, 5'-YTA[AT](4)TAR-3', found in numerous muscle-specific, growth factor- and stress-induced genes. Mediates cellular functions not only in skeletal and cardiac muscle development, but also in neuronal differentiation and survival. Plays diverse roles in the control of cell growth, survival and apoptosis via p38 MAPK signaling in muscle-specific and/or growth factor-related transcription. Plays a critical role in the regulation of neuronal apoptosis. -
序列相似性
Belongs to the MEF2 family.
Contains 1 MADS-box domain.
Contains 1 Mef2-type DNA-binding domain. -
发展阶段
Present in myotubes and also in undifferentiated myoblasts. -
结构域
The beta domain, missing in a number of isoforms, is required for enhancement of transcriptional activity. -
翻译后修饰
Phosphorylated on Ser-444 by CDK5 is required for Lys-439 sumoylation and inhibits transcriptional activity. In neurons, enhanced CDK5 activity induced by neurotoxins promotes caspase 3-mediated cleavage leading to neuron apoptosis. Phsophorylation on Ser-180 can be enhanced by EGF.
Acetylated on Lys-439 by CREBBP. Deacetylated by SIRT1.
Sumoylated on Lys-439 by SUMO2 but not SUMO1; which inhibits transcriptional activity and myogenic activity. Desumoylated by SENP3.
Proteolytically cleaved in cerebellar granule neurons on several sites by caspase 7 following neurotoxicity. Preferentially cleaves the CDK5-mediated hyperphosphorylated form which leads to neuron apoptosis and transcriptional inactivation. -
细胞定位
Nucleus. Translocated by HDAC4 to nuclear dots. - Information by UniProt
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数据库链接
- Entrez Gene: 4209 Human
- Entrez Gene: 17261 Mouse
- Entrez Gene: 81518 Rat
- Omim: 600663 Human
- SwissProt: Q14814 Human
- SwissProt: Q63943 Mouse
- SwissProt: O89038 Rat
- Unigene: 314327 Human
see all -
别名
- DKFZp686I1536 antibody
- MADS box transcription factor 2 polypeptide D antibody
- Mef2d antibody
see all
图片
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All lanes : Anti-MEF2D antibody [EPR24993-10] (ab282731) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : mef2d CRISPR-Cas9 edited HeLa cell lysate
Lane 3 : U-2 OS cell lysate
Lane 4 : NIH/3T3 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 56 kDa
Observed band size: 67 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-MEF2D antibody [EPR24993-10] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab282731 was shown to bind specifically to MEF2D. A band was observed at 67 kDa in wild-type HeLa cell lysates with no signal observed at this size in mef2d CRISPR-Cas9 edited cell line ab281636. The band observed in the CRISPR-Cas9 edited lysate lane below 67 kDa is likely to represent a truncated form of MEF2D. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and mef2d CRISPR-Cas9 edited HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) 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-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labelling MEF2D with ab282731 at 1/2000 (0.286 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on mouse testis (PMID: 24694307). The section was incubated with ab282731 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized THP-1 cells labelling MEF2D with ab282731 at 1/50 (11.42 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green) Confocal image showing mainly nuclear staining in THP-1 cells is observed.
ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized THP-1 (Human monocytic leukemia monocyte) cells labelling MEF2D with ab282731 at 1/500 dilution (0.1 ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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All lanes : Anti-MEF2D antibody [EPR24993-10] (ab282731) at 1/1000 dilution
Lane 1 : THP-1 (Human monocytic leukemia monocyte) whole cell lysates
Lane 2 : SW480 (human colorectal adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : DLD-1 (human colon epithelial) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 56 kDa
Observed band size: 70, 40 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Low expression: SW480/ DLD-1 (PMID: 27364559).
Exposure time: 147 seconds
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All lanes : Anti-MEF2D antibody [EPR24993-10] (ab282731) at 1/1000 dilution
Lane 1 : RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 2 : 2.4G2 (rat B cell lymphoma B lymphocyte) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 56 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Lysates were made freshly and used in WB test immediately to minimize protein degradation.
Exposure time: 147 seconds
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Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling MEF2D with ab282731 at 1/2000 (0.286 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on mouse cererbum (PMID: 28738418). The section was incubated with ab282731 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
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All lanes : Anti-MEF2D antibody [EPR24993-10] (ab282731) at 1/1000 dilution
Lane 1 : His-tagged human MEF2D recombinant protein
Lane 2 : Myc/DDDDK-tagged human MEF2A recombinant protein
Lane 3 : Myc/DDDDK-tagged human MEF2B recombinant protein
Lane 4 : Myc/DDDDK-tagged human MEF2C recombinant protein
Lysates/proteins at 0.01 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 56 kDa
Observed band size: 31 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
His-tagged human MEF2D recombinant protein (aa54-328) 10ng
Myc/DDDDK-tagged human MEF2A recombinant protein 10ng
Myc/DDDDK-tagged human MEF2B recombinant protein 10ng
Myc/DDDDK-tagged human MEF2C recombinant protein 10ng
Exposure time: 10 seconds
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Immunohistochemical analysis of paraffin-embedded Rat colon tissue labelling MEF2D with ab282731 at 1/2000 (0.286 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on rat colon. The section was incubated with ab282731 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
-
Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labelling MEF2D with ab282731 at 1/2000 (0.286 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on rat cererbum (PMID: 28738418). The section was incubated with ab282731 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
-
Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 cells labelling MEF2D with ab282731 at 1/50 (11.42 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green) Confocal image showing mainly nuclear staining in RAW 264.7 cells is observed.
ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling MEF2D with ab282731 at 1/500 dilution (0.1 ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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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SDS download
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Datasheet download
Certificate of Compliance
文献 (0)
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