重组Anti-PDHA1抗体[EPR11098] (ab168379)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR11098] to PDHA1
- Suitable for: Flow Cyt (Intra), WB, ICC/IF, IHC-P, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-PDHA1抗体[EPR11098]
参阅全部 PDHA1 一抗 -
描述
兔单克隆抗体[EPR11098] to PDHA1 -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), WB, ICC/IF, IHC-P, IPmore 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
- Human fetal kidney, A549, Jurkat, HepG2 and HeLa lysates; Human kidney and skeletal muscle tissues; HepG2 cells; permeabilized Jurkat cells; HT-29; Mouse kidney; Rat kidney.
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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
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR11098 -
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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KO cell lysates
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Positive Controls
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab168379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) |
1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
1/1000 - 1/5000. Predicted molecular weight: 43 kDa.
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ICC/IF |
1/100 - 1/500.
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IP |
1/10 - 1/100.
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说明 |
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Flow Cyt (Intra)
1/10 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/1000 - 1/5000. Predicted molecular weight: 43 kDa. |
ICC/IF
1/100 - 1/500. |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IP
1/10 - 1/100. |
靶标
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功能
The pyruvate dehydrogenase complex catalyzes the overall conversion of pyruvate to acetyl-CoA and CO(2). It contains multiple copies of three enzymatic components: pyruvate dehydrogenase (E1), dihydrolipoamide acetyltransferase (E2) and lipoamide dehydrogenase (E3). -
组织特异性
Ubiquitous. -
疾病相关
Defects in PDHA1 are a cause of pyruvate decarboxylase E1 component deficiency (PDHE1 deficiency) [MIM:312170]. PDHE1 deficiency is the most common enzyme defect in patients with primary lactic acidosis. It is associated with variable clinical phenotypes ranging from neonatal death to prolonged survival complicated by developmental delay, seizures, ataxia, apnea, and in some cases to an X-linked form of Leigh syndrome (X-LS).
Defects in PDHA1 are the cause of X-linked Leigh syndrome (X-LS) [MIM:308930]. X-LS is an early-onset progressive neurodegenerative disorder with a characteristic neuropathology consisting of focal, bilateral lesions in one or more areas of the central nervous system, including the brainstem, thalamus, basal ganglia, cerebellum, and spinal cord. The lesions are areas of demyelination, gliosis, necrosis, spongiosis, or capillary proliferation. Clinical symptoms depend on which areas of the central nervous system are involved. The most common underlying cause is a defect in oxidative phosphorylation. LS may be a feature of a deficiency of any of the mitochondrial respiratory chain complexes. -
细胞定位
Mitochondrion matrix. - Information by UniProt
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数据库链接
- Entrez Gene: 5160 Human
- Entrez Gene: 18597 Mouse
- Entrez Gene: 29554 Rat
- Omim: 300502 Human
- SwissProt: P08559 Human
- SwissProt: P35486 Mouse
- SwissProt: P26284 Rat
- Unigene: 530331 Human
see all -
别名
- ODPA_HUMAN antibody
- PDH antibody
- PDHA antibody
see all
图片
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All lanes : Anti-PDHA1 antibody [EPR11098] (ab168379) at 1/1000 dilution
Lane 1 : Wild-type A549 whole cell lysate
Lane 2 : PDHA1 knockout A549 whole cell lysate
Lane 3 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 43 kDa
Observed band size: 43 kDaLanes 1 - 3: Merged signal (red and green). Green - ab168379 observed at 43 kDa. Red - loading control, ab130007, observed at 130 kDa.
ab168379 was shown to recognize PDHA1 in wild-type A549 cells as signal was lost at the expected MW in PDHA1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and PDHA1 knockout samples were subjected to SDS-PAGE. Ab168379 and ab130007 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-PDHA1 antibody [EPR11098] (ab168379) at 1/2000 dilution
Lane 1 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates
Lane 2 : Mouse brain lysates
Lane 3 : Rat brain lysates
Lane 4 : Mouse kidney lysates
Lane 5 : Rat kidney lysates
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: 43 kDa
Observed band size: 43 kDaBlocking and diluting buffer: 5% NFDM/TBST
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Immunocytochemistry/ Immunofluorescence analysis of HT-29 (Human colorectal adenocarcinoma epithelial cell) cells labeling PDHA1 with Purified ab168379 at 1:100 dilution (3.5 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). ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ab168379 (purified) at 1:20 dilution (2ug) immunoprecipitating PDHA1 in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug
Lane 2 (+): ab168379 & HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab168379 in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
Intracellular Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling PDHA1 with purified ab168379 at 1/40 dilution (10 ug/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilized with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue sections labeling PDHA1 with Purified ab168379 at 1:200 dilution (1.76 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, pH 9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) 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) analysis of human thyroid carcinoma tissue sections labeling PDHA1 with Purified ab168379 at 1:200 dilution (1.76 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, pH 9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) 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) analysis of mouse kidney tissue sections labeling PDHA1 with Purified ab168379 at 1:200 dilution (1.76 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, pH 9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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Immunocytochemistry/Immunofluorescence analysis Jurkat (human acute T cell leukemia) labelling PDHA1 with purified ab168379 at 1/500. Cells were fixed with 100% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).
Control: PBS only
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All lanes : Anti-PDHA1 antibody [EPR11098] (ab168379) at 1/1000 dilution (unpurified)
Lane 1 : Human fetal kidney lysate
Lane 2 : A549 lysate
Lane 3 : Jurkat lysate
Lane 4 : HepG2 lysate
Lane 5 : HeLa lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-rabbit HRP at 1/2000 dilution
Predicted band size: 43 kDa -
Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling PDHA1 with unpurified ab168379 at 1/100 dilution. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling PDHA1 with unpurified ab168379 at 1/100 dilution. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.
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Immunofluorescent analysis of HepG2 cells labeling PDHA1 with unpurified ab168379 at 1/100 dilution.
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Intracellular flow cytometric analysis of permeabilized Jurkat cells labeling PDHA1 (red) with unpurified ab168379 at 1/10 dilution, or a rabbit IgG (negative) (green).
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Detection of PDHA1 by Western Blot of Immunprecipitate. 293T cell lysate immunoprecipitated using unpurified ab168379 at 1/10 dilution; HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (22)
ab168379 被引用在 22 文献中.
- Pin F et al. The Mitochondria-Targeting Agent MitoQ Improves Muscle Atrophy, Weakness and Oxidative Metabolism in C26 Tumor-Bearing Mice. Front Cell Dev Biol 10:861622 (2022). PubMed: 35392166
- Lan T et al. Effects of MICU1-Mediated Mitochondrial Calcium Uptake on Energy Metabolism and Quality of Vitrified-Thawed Mouse Metaphase II Oocytes. Int J Mol Sci 23:N/A (2022). PubMed: 35955764
- Meng L et al. Dysregulation of the Sirt5/IDH2 axis contributes to sunitinib resistance in human renal cancer cells. FEBS Open Bio 11:921-931 (2021). PubMed: 33455080
- Shi W et al. Downregulation of GLUT3 impairs STYK1/NOK-mediated metabolic reprogramming and proliferation in NIH-3T3 cells. Oncol Lett 22:527 (2021). PubMed: 34055092
- Tardo-Dino PE et al. Effect of heat acclimation on metabolic adaptations induced by endurance training in soleus rat muscle. Physiol Rep 9:e14686 (2021). PubMed: 34405575