Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Pyruvate Dehydrogenase E1-alpha subunit antibody [EPR11099] (ab155096)

Overview

  • Product name

    Anti-Pyruvate Dehydrogenase E1-alpha subunit antibody [EPR11099]
    See all Pyruvate Dehydrogenase E1-alpha subunit primary antibodies
  • Description

    Rabbit monoclonal [EPR11099] to Pyruvate Dehydrogenase E1-alpha subunit
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide, corresponding to residues in Human Pyruvate Dehydrogenase E1-alpha subunit (UniProt: P08559).

  • Positive control

    • HepG2, 293T, HeLa, and Jurkat whole cell lysate (ab7899); Human thyroid carcinoma tissue; HeLa cells.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab155096 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 1/1000 - 1/2000. Predicted molecular weight: 43 kDa.
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF 1/250 - 1/500.
IP 1/10 - 1/100.
Flow Cyt 1/1000 - 1/10000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

Target

  • 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).
  • Tissue specificity

    Ubiquitous.
  • Involvement in disease

    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.
  • Cellular localization

    Mitochondrion matrix.
  • Information by UniProt
  • Database links

  • Alternative names

    • ODPA_HUMAN antibody
    • PDH antibody
    • PDHA antibody
    • PDHA1 antibody
    • PDHCE1A antibody
    • PDHE1 A type I antibody
    • PDHE1-A type I antibody
    • PHE1A antibody
    • Pyruvate Dehydrogenase (lipoamide) alpha 1 antibody
    • Pyruvate dehydrogenase complex, E1 alpha polypeptide 1 antibody
    • Pyruvate Dehydrogenase E1 alpha antibody
    • Pyruvate dehydrogenase E1 component subunit alpha, somatic form, mitochondrial antibody
    see all

Images

  • All lanes : Anti-Pyruvate Dehydrogenase E1-alpha subunit antibody [EPR11099] (ab155096) at 1/1000 dilution

    Lane 1 : Wild-type HeLa whole cell lysate
    Lane 2 : PDHA1 knockout HeLa whole cell lysate
    Lane 3 : HEK-293 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 43 kDa
    Observed band size: 43 kDa



    Lanes 1 - 3: Merged signal (red and green). Green - ab155096 observed at 43 kDa. Red - loading control, ab130007, observed at 130 kDa.

    ab155096 was shown to recognize PDHA1 in wild-type HeLa 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. The membrane was blocked with 3% Milk. Ab155096 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.

  • All lanes : Anti-Pyruvate Dehydrogenase E1-alpha subunit antibody [EPR11099] (ab155096) at 1/1000 dilution

    Lane 1 : HepG2 cell lysate
    Lane 2 : 293T cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : Jurkat cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit HRP at 1/2000 dilution

    Predicted band size: 43 kDa

  • Overlay histogram showing HeLa cells stained with ab155096 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab155096, 1/10000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
  • Immunohistochemical analysis of paraffin-embedded Human thyroid carcinoma tissue labeling Pyruvate Dehydrogenase E1-alpha subunit with ab155096 at 1/50 dilution.
  • Immunofluorescent analysis of HeLa cells labeling Pyruvate Dehydrogenase E1-alpha subunit with ab155096 at 1/250 dilution.

References

This product has been referenced in:

  • Bickert A  et al. Inactivation of ceramide synthase 2 catalytic activity in mice affects transcription of genes involved in lipid metabolism and cell division. Biochim Biophys Acta 1863:734-749 (2018). WB . Read more (PubMed: 29653252) »
  • Malty RH  et al. A Map of Human Mitochondrial Protein Interactions Linked to Neurodegeneration Reveals New Mechanisms of Redox Homeostasis and NF-?B Signaling. Cell Syst 5:564-577.e12 (2017). Read more (PubMed: 29128334) »
See all 3 Publications for this product

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