Anti-Pyruvate Dehydrogenase E2 antibody [15D3G9C11] (ab110332)
![Western blot - Anti-Pyruvate Dehydrogenase E2 antibody [15D3G9C11] (ab110332)](https://www.abcam.com/ps/products/110/ab110332/Images/ab110332-365487-anti-pyruvate-dehydrogenase-e2-antibody-15d3g9c11-western-blot-knockout-hap1-dlat-hela-hl60-human-lysates.jpg "Western blot - Anti-Pyruvate Dehydrogenase E2 antibody [15D3G9C11] (ab110332)")
Key features and details
- Mouse monoclonal [15D3G9C11] to Pyruvate Dehydrogenase E2
- Suitable for: WB, ICC/IF, Flow Cyt, IHC-P
- Knockout validated
- Reacts with: Human
- Isotype: IgG1
Overview
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Product name
Anti-Pyruvate Dehydrogenase E2 antibody [15D3G9C11]
See all Pyruvate Dehydrogenase E2 primary antibodies -
Description
Mouse monoclonal [15D3G9C11] to Pyruvate Dehydrogenase E2 -
Host species
Mouse -
Tested applications
Suitable for: WB, ICC/IF, Flow Cyt, IHC-Pmore details -
Species reactivity
Reacts with: Human -
Immunogen
Full length protein. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- Isolated mitochondria from Human heart; Normal Human embryonic lung fibroblasts (strain MRC5); Human cerebellum tissue; HL60 cells.
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General notes
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Product was previously marketed under the MitoSciences sub-brand.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.5
Preservative: 0.02% Sodium azide
Constituent: HEPES buffered saline -
Concentration information loading... -
Purity
IgG fraction -
Purification notes
ab110332 was produced in vitro using hybridomas grown in serum-free medium, and then purified by biochemical fractionation. -
Clonality
Monoclonal -
Clone number
15D3G9C11 -
Isotype
IgG1 -
Light chain type
kappa -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab110332 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
Use a concentration of 0.5 µg/ml. Predicted molecular weight: 69 kDa.
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| ICC/IF |
Use a concentration of 0.2 - 0.5 µg/ml.
(heat-induced antigen-retrieval improvessignal). |
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| Flow Cyt |
Use a concentration of 1 µg/ml.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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| IHC-P |
1/100. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
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| Notes |
|---|
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WB
Use a concentration of 0.5 µg/ml. Predicted molecular weight: 69 kDa. |
|
ICC/IF
Use a concentration of 0.2 - 0.5 µg/ml. (heat-induced antigen-retrieval improvessignal). |
|
Flow Cyt
Use a concentration of 1 µg/ml. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
|
IHC-P
1/100. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol. |
Target
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Function
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). -
Involvement in disease
Note=Primary biliary cirrhosis is a chronic, progressive cholestatic liver disease characterized by the presence of antimitochondrial autoantibodies in patients' serum. It manifests with inflammatory obliteration of intra-hepatic bile duct, leading to liver cell damage and cirrhosis. Patients with primary biliary cirrhosis show autoantibodies against the E2 component of pyruvate dehydrogenase complex.
Defects in DLAT are the cause of pyruvate dehydrogenase E2 deficiency (PDHE2 deficiency) [MIM:245348]; also known as lactic acidemia due to defect of E2 lipoyl transacetylase of the pyruvate dehydrogenase complex. Pyruvate dehydrogenase (PDH) deficiency is a major cause of primary lactic acidosis and neurological dysfunction in infancy and early childhood. In this form of PDH deficiency episodic dystonia is the major neurological manifestation, with other more common features of pyruvate dehydrogenase deficiency, such as hypotonia and ataxia, being less prominent. -
Sequence similarities
Belongs to the 2-oxoacid dehydrogenase family.
Contains 2 lipoyl-binding domains. -
Cellular localization
Mitochondrion matrix. - Information by UniProt
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Database links
- Entrez Gene: 1737 Human
- Omim: 608770 Human
- SwissProt: P10515 Human
- Unigene: 335551 Human
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Alternative names
- 70 kDa mitochondrial autoantigen of primary biliary cirrhosis antibody
- Dihydrolipoamide acetyltransferase component of pyruvate dehydrogenase complex antibody
- Dihydrolipoamide S Acetyltransferase antibody
see all
Images
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Western blot - Anti-Pyruvate Dehydrogenase E2 antibody [15D3G9C11] (ab110332)All lanes : Anti-Pyruvate Dehydrogenase E2 antibody [15D3G9C11] (ab110332) at 0.5 µg
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : DLAT knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : HL-60 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 69 kDaLanes 1 - 4: Merged signal (red and green). Green - ab110332 observed at 72 kDa. Red - loading control, ab181602, observed at 38 kDa.
ab110332 was shown to specifically react with in wild-type HAP1 cells as signal was lost in DLAT knockout cells. Wild-type and DLAT knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab110332 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 0.5 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 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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![Immunocytochemistry/ Immunofluorescence - Anti-Pyruvate Dehydrogenase E2 antibody [15D3G9C11] (ab110332)](https://www.abcam.com/ps/products/110/ab110332/Images/ab110332-123648-anti-pyruvate-dehydrogenase-e2-antibody-15d3g9c11-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Pyruvate Dehydrogenase E2 antibody [15D3G9C11] (ab110332)Immunocytochemistry analysis using ab110332 at 1µg/ml staining Pyruvate Dehydrogenase E2 in cultured, normal Human embryonic lung fibroblasts (strain MRC5) and an AlexaFluor® 488 goat anti-mouse IgG1 secondary antibody (2 ug/ml). -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pyruvate Dehydrogenase E2 antibody [15D3G9C11] (ab110332)](https://www.abcam.com/ps/products/110/ab110332/Images/ab110332-123647-anti-pyruvate-dehydrogenase-e2-antibody-15d3g9c11-immunohistochemistry.gif)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pyruvate Dehydrogenase E2 antibody [15D3G9C11] (ab110332)Immunohistological analysis using ab110332 at 1/100 dilution staining Pyruvate Dehydrogenase E2 in Human cerebellum tissue (Formalin-fixed, Paraffin-embedded). -
![Flow Cytometry - Anti-Pyruvate Dehydrogenase E2 antibody [15D3G9C11] (ab110332)](https://www.abcam.com/ps/products/110/ab110332/Images/ab110332-413043-AntiPyruvate-Dehydrogenase-E2-antibody.jpg)
Flow Cytometry - Anti-Pyruvate Dehydrogenase E2 antibody [15D3G9C11] (ab110332)ICC/IF image of ab109866 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab109866 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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![Western blot - Anti-Pyruvate Dehydrogenase E2 antibody [15D3G9C11] (ab110332)](https://www.abcam.com/ps/products/110/ab110332/Images/ab110332-123649-anti-pyruvate-dehydrogenase-e2-antibody-15d3g9c11-western-blot.jpg)
Western blot - Anti-Pyruvate Dehydrogenase E2 antibody [15D3G9C11] (ab110332)Anti-Pyruvate Dehydrogenase E2 antibody [15D3G9C11] (ab110332) at 0.5 µg/ml + Isolated mitochondria from Human heart at 5 µg
Predicted band size: 69 kDa -
![Immunocytochemistry/ Immunofluorescence - Anti-Pyruvate Dehydrogenase E2 antibody [15D3G9C11] (ab110332)](https://www.abcam.com/ps/products/110/ab110332/Images/ab110332-413042-AntiPyruvate-Dehydrogenase-E2-antibody.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Pyruvate Dehydrogenase E2 antibody [15D3G9C11] (ab110332)ICC/IF image of ab109866 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab109866 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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![Flow Cytometry - Anti-Pyruvate Dehydrogenase E2 antibody [15D3G9C11] (ab110332)](https://www.abcam.com/ps/products/110/ab110332/Images/ab110332-123646-anti-pyruvate-dehydrogenase-e2-antibody-15d3g9c11-flow-cytometry.gif)
Flow Cytometry - Anti-Pyruvate Dehydrogenase E2 antibody [15D3G9C11] (ab110332)Flow cytometric analysis using ab110332 at 1µg/ml staining Pyruvate Dehydrogenase E2 in HL60 cells (blue). Isotype control antibody (red).
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (8)
ab110332 has been referenced in 8 publications.
- An YA et al. Dysregulation of Amyloid Precursor Protein Impairs Adipose Tissue Mitochondrial Function and Promotes Obesity. Nat Metab 1:1243-1257 (2019). PubMed: 31984308
- Matsuda S et al. Nuclear pyruvate kinase M2 complex serves as a transcriptional coactivator of arylhydrocarbon receptor. Nucleic Acids Res 44:636-47 (2016). PubMed: 26405201
- Sloan JL et al. Exome sequencing identifies ACSF3 as a cause of combined malonic and methylmalonic aciduria. Nat Genet : (2011). WB ; Human . PubMed: 21841779
- Feng D et al. Down-regulation of mitochondrial acyl carrier protein in mammalian cells compromises protein lipoylation and respiratory complex I and results in cell death. J Biol Chem 284:11436-45 (2009). PubMed: 19221180
- Ridout CK et al. Somatic mosaicism for a PDHA1 mutation in a female with pyruvate dehydrogenase deficiency. Hum Genet 124:187-93 (2008). PubMed: 18709504
- Han Z et al. A combined therapeutic approach for pyruvate dehydrogenase deficiency using self-complementary adeno-associated virus serotype-specific vectors and dichloroacetate. Mol Genet Metab 93:381-7 (2008). PubMed: 18206410
- Stiburek L et al. Knockdown of human Oxa1l impairs the biogenesis of F1Fo-ATP synthase and NADH:ubiquinone oxidoreductase. J Mol Biol 374:506-16 (2007). PubMed: 17936786
- Koukourakis MI et al. Pyruvate dehydrogenase and pyruvate dehydrogenase kinase expression in non small cell lung cancer and tumor-associated stroma. Neoplasia 7:1-6 (2005). PubMed: 15736311