Anti-Pyruvate Dehydrogenase E1-alpha subunit antibody [8D10E6] (ab110334)

Overview

  • Product name

    Anti-Pyruvate Dehydrogenase E1-alpha subunit antibody [8D10E6]
    See all Pyruvate Dehydrogenase E1-alpha subunit primary antibodies
  • Description

    Mouse monoclonal [8D10E6] to Pyruvate Dehydrogenase E1-alpha subunit
  • Host species

    Mouse
  • Tested applications

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

    Reacts with: Mouse, Rat, Cow, Human, Caenorhabditis elegans, Drosophila melanogaster, Zebrafish
  • Immunogen

    Full length native protein (purified). This information is considered to be commercially sensitive.

  • Positive control

    • WB: Isolated mitochondria from human, bovine, rat and mouse heart. HepG2 (human liver hepatocellular carcinoma cell line) cell lysate. FACS: HeLa (human epithelial cell line from cervix adenocarcinoma) and HL-60 (human promyelocytic leukemia cell line) cells.
  • General notes

    This antibody clone is manufactured by Abcam.

    Product was previously marketed under the MitoSciences sub-brand.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab110334 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
IHC-Fr Use at an assay dependent concentration. PubMed: 25223649
WB Use a concentration of 0.5 - 1 µg/ml. Predicted molecular weight: 43 kDa.
ICC/IF Use a concentration of 1 µg/ml. (heat-induced antigen-retrieval improves signal)
Flow Cyt Use a concentration of 1 µg/ml.

ab170190 - Mouse monoclonal IgG1, 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 antibody
    • Pyruvate dehydrogenase E1 component subunit alpha, somatic form, mitochondrial antibody
    see all

Images

  • All lanes : Anti-Pyruvate Dehydrogenase E1-alpha subunit antibody [8D10E6] (ab110334) at 0.5 µg/ml

    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 - ab110334 observed at 43 kDa. Red - loading control, ab52866, observed at 50 kDa.

    ab110334 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. Ab110334 and ab52866 (Rabbit anti alpha Tubulin loading control) were incubated overnight at 4°C at 0.5 ug/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • All lanes : Anti-Pyruvate Dehydrogenase E1-alpha subunit antibody [8D10E6] (ab110334) at 1 µg/ml

    Lane 1 : Isolated mitochondria from human heart at 5 µg
    Lane 2 : Isolated mitochondria from bovine heart at 1 µg
    Lane 3 : Isolated mitochondria from rat heart at 10 µg
    Lane 4 : Isolated mitochondria from mouse heart at 10 µg
    Lane 5 : HepG2 cell lysate at 20 µg

    Predicted band size: 43 kDa

  • Immunocytochemistry analysis using ab110334 at 1 µg/ml staining Pyruvate Dehydrogenase E1-alpha subunit in HeLa (human epithelial cell line from cervix adenocarcinoma) cells (4% paraformaldehyde fixed and Triton X-100 permeabilized).
    The secondary antibody was (green) Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution. DAPI was used to stain the cell nuclei (blue).

  • Flow cytometric analysis using ab110334 at 1 µg/ml staining Pyruvate Dehydrogenase E1-alpha subunit in HL-60 (human promyelocytic leukemia cell line) cells (blue). Isotype control antibody (red).

References

This product has been referenced in:

  • Zhuang Y  et al. The novel function of tumor protein D54 in regulating pyruvate dehydrogenase and metformin cytotoxicity in breast cancer. Cancer Metab 7:1 (2019). Read more (PubMed: 30697423) »
  • Xu L  et al. The SIRT2/cMYC Pathway Inhibits Peroxidation-Related Apoptosis In Cholangiocarcinoma Through Metabolic Reprogramming. Neoplasia 21:429-441 (2019). Read more (PubMed: 30933885) »
See all 36 Publications for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

Application
Western blot
Sample
Sheep Tissue lysate - whole (Skeletal Muscle)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
50 µg
Specification
Skeletal Muscle
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Alex Pendleton

Verified customer

Submitted Nov 14 2017

Application
Western blot
Sample
Rat Tissue lysate - whole (Retina)
Gel Running Conditions
Reduced Denaturing (12% gel)
Loading amount
20 µg
Specification
Retina
Blocking step
Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 01 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (HEK cells)
Gel Running Conditions
Reduced Denaturing (12% gel)
Loading amount
20 µg
Specification
HEK cells
Blocking step
Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 03 2017

Application
Western blot
Sample
Mouse Tissue lysate - whole (Liver, retina)
Gel Running Conditions
Reduced Denaturing (12% gel)
Loading amount
20 µg
Specification
Liver, retina
Blocking step
Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 03 2017

Application
Western blot
Sample
Zebrafish Tissue lysate - whole (Retina)
Gel Running Conditions
Reduced Denaturing (12% gel)
Loading amount
20 µg
Specification
Retina
Blocking step
Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 03 2017

Application
Western blot
Sample
Euglena Cell lysate - whole cell (Euglena gracilis SM-ZK)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
15 µg
Treatment
silencing of pyruvate dehydrogenase E1-alpha subunit or not
Specification
Euglena gracilis SM-ZK
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jul 07 2016

Answer

Thank you for contacting us.

We unfortunately do not provide free samples, this is why we have 100% Abtrial offer. If this antibody does not work you will still be getting a free antibody with Abtrial code.

Please let me know if you are interested in getting this code.

Thanks!

Read More

Answer

Thank you for contacting us.

This product ab110330 was tested with Mouse, Rat, Cow and Human samples only; horse was never tried. We can however predict, it will be fine for horse samples as well because the horse protein (http://www.uniprot.org/uniprot/F6WQM7) is quite homologous to the human protein (http://www.uniprot.org/uniprot/P08559).

In case you are interested in trying this antibody then I would encourage you to participate in our 100% Abtrial discount programme - a free product can be claimed in exchange of review for this antibody. Please contact me for code if you accept our offer.

More details about the offer can be found at the following link;

https://www.abcam.com/index.html?pageconfig=resource&rid=11998&viapagetrap=collaborationdiscount


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thank you for contacting us. I am sorry that these antibodies are no longer working properly. I will be happy to help you resolve this issue if you can please provide some additional details about your experiment?

1.) What application were you using these antibodies in (WB, IHC-P, etc)?

2.) What species, tissue type, or cell line were you testing? How were the samples prepared (type of buffer used for lysis, fixation, permeabilization, embedding method, etc)?

3.) What blocking agent was used and at what concentration?

4.) What were the primary and secondary antibody dilutions and incubation times?

5.) Could you please provide a description of the problem (no signal, high background, etc) and / or an image of your results?

6.) What was your original order or PO number?

I will be happy to help you further once I have this additional information.

Read More

Answer

Thank you for contacting us.

I found the original order for these ordered on July 28 2011. Since these antibodies are recommended to be stored at 4C, they only have a shelf-life of roughly 1 yr. If the end-user is seeing decreased signal and the antibodies worked well in the past they may have to increase the working dilution and/or incubation time.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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1-10 of 11 Abreviews or Q&A

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