重组Anti-IDH2抗体[EPR7577] (ab131263)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR7577] to IDH2
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-IDH2抗体[EPR7577]
参阅全部 IDH2 一抗 -
描述
兔单克隆抗体[EPR7577] to IDH2 -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- Wild-type A549, Wild-type Jurkat, MOLT-4, K562, U-87 MG, and HepG2 cell lysates.
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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 -20°C. Stable for 12 months at -20°C. -
解离常数(KD)
KD = 4.70 x 10 -11 M Learn more about KD -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR7577 -
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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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab131263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) |
1/100 - 1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
1/1000 - 1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 50 kDa).
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF | (1) |
1/1000.
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说明 |
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Flow Cyt (Intra)
1/100 - 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/1000 - 1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 50 kDa). |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
1/1000. |
靶标
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功能
Plays a role in intermediary metabolism and energy production. It may tightly associate or interact with the pyruvate dehydrogenase complex. -
疾病相关
D-2-hydroxyglutaric aciduria 2
Glioma
enetic variations are associated with cartilaginous tumors such as enchondroma or chondrosarcoma. -
序列相似性
Belongs to the isocitrate and isopropylmalate dehydrogenases family. -
翻译后修饰
Acetylation at Lys-413 dramatically reduces catalytic activity. Deacetylated by SIRT3. -
细胞定位
Mitochondrion. - Information by UniProt
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数据库链接
- Entrez Gene: 3418 Human
- Entrez Gene: 269951 Mouse
- Entrez Gene: 361596 Rat
- Omim: 147650 Human
- SwissProt: P48735 Human
- SwissProt: P54071 Mouse
- SwissProt: P56574 Rat
- Unigene: 596461 Human
see all -
别名
- D2HGA2 antibody
- ICD-M antibody
- IDH antibody
see all
图片
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All lanes : Anti-IDH2 antibody [EPR7577] (ab131263) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : IDH2 knockout A549 cell lysate
Lane 3 : Wild-type Jurkat cell lysate
Lane 4 : IDH2 knockout Jurkat ab282331 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 50 kDa
Observed band size: 51 kDa why is the actual band size different from the predicted?Western blot: Anti-IDH2 antibody [EPR7577] (ab131263) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab131263 was shown to bind specifically to IDH2. A band was observed at 51 kDa in wild-type A549 cell lysates with no signal observed at this size in IDH2 knockout cell line. To generate this image, wild-type and IDH2 knockout A549 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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All lanes : Anti-IDH2 antibody [EPR7577] (ab131263) at 1/1000 dilution
Lane 1 : Wild-type Jurkat cell lysate
Lane 2 : IDH2 knockout Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 50 kDa
Observed band size: 48 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-IDH2 antibody [EPR7577] 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, ab131263 was shown to bind specifically to IDH2. A band was observed at 48 kDa in wild-type Jurkat cell lysates with no signal observed at this size in IDH2 knockout cell line ab282331 (knockout cell lysate ab283148). To generate this image, wild-type and IDH2 knockout Jurkat 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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Lanes 1-7 : Anti-IDH2 antibody [EPR7577] (ab131263) at 1/5000 dilution (Purified)
Lanes 8-9 : Anti-IDH2 antibody [EPR7577] (ab131263) at 1/5000 dilution
Lane 1 : MOLT-4 (Human lymphoblastic leukemia T lymphoblast) whole cell lysate
Lane 2 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate
Lane 3 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysate
Lane 4 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 5 : Mouse liver lysate
Lane 6 : Rat liver lysate
Lane 7 : Mouse kidney lysate
Lane 8 : Rat kidney lysate
Lane 9 : Rat stomach lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 50 kDa -
Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labelling IDH2 with Purified ab131263 at 1:20 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling IDH2 with Purified ab131263 at 1:1000 dilution (0.2 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH2 antibody [EPR7577] (ab131263)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue sections labeling IDH2 with Purified ab131263 at 1:100 (2.11 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH2 antibody [EPR7577] (ab131263)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse spleen tissue sections labeling IDH2 with Purified ab131263 at 1:100 (2.11 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH2 antibody [EPR7577] (ab131263)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat spleen tissue sections labeling IDH2 with Purified ab131263 at 1:100 (2.11 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: IDH2 knockout HAP1 cell lysate (20 µg)
Lane 3: K562 cell lysate (20 µg)
Lane 4: HepG2 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab131263 observed at 48 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab131263 was shown to recognize IDH2 when IDH2 knockout samples were used, along with additional cross-reactive bands. Wild-type and IDH2 knockout samples were subjected to SDS-PAGE. ab131263 and ab8245 (loading control to GAPDH) were both diluted 1/10 000 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 h at room temperature before imaging.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (17)
ab131263 被引用在 17 文献中.
- Li J et al. Energy Metabolic Disorder of Astrocytes May Be an Inducer of Migraine Attack. Brain Sci 12:N/A (2022). PubMed: 35884650
- Wang Z et al. Impaired tricarboxylic acid cycle flux and mitochondrial aerobic respiration during isoproterenol induced myocardial ischemia is rescued by bilobalide. J Pharm Anal 11:764-775 (2021). PubMed: 35028182
- Yu Y et al. SUMOylation enhances the activity of IDH2 under oxidative stress. Biochem Biophys Res Commun 532:591-597 (2020). PubMed: 32900482
- Ji D et al. Effects of inflammatory and anti-inflammatory environments on the macrophage mitochondrial function. Sci Rep 10:20324 (2020). PubMed: 33230189
- Seok J et al. The oncometabolite d-2-hydroxyglutarate induces angiogenic activity through the vascular endothelial growth factor receptor 2 signaling pathway. Int J Oncol 54:753-763 (2019). PubMed: 30483760