Recombinant
RabMAb

Recombinant Anti-Pyruvate Dehydrogenase E1-alpha subunit (phospho S293) antibody [EPR12200] (ab177461)

Overview

  • Product name
    Anti-Pyruvate Dehydrogenase E1-alpha subunit (phospho S293) antibody [EPR12200]
    See all Pyruvate Dehydrogenase E1-alpha subunit primary antibodies
  • Description
    Rabbit monoclonal [EPR12200] to Pyruvate Dehydrogenase E1-alpha subunit (phospho S293)
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, IP, ELISAmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    within Human Pyruvate Dehydrogenase E1-alpha subunit aa 250-350 (Cysteine residue). The exact sequence is proprietary.
    Database link: P08559

  • Positive control
    • 293T cell lysates; Human breast and colon tissue; HT-29; Mouse kidney; Mouse colon; Rat kidney.
  • 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab177461 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 - 1/5000. Predicted molecular weight: 40 kDa.
IHC-P 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

For unpurified use at 1/50 - 1/100. 

IP 1/10 - 1/100.
ELISA Use at an assay dependent concentration.

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 (phospho S293) antibody [EPR12200] (ab177461) at 1/2000 dilution (purified)

    Lane 1 : Rat kidney lysates
    Lane 2 : Rat kidney lysates.Then the membrane was incubated with phosphatase.

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 40 kDa



    Blocking and diluting buffer : 5% NFDM/TBST
  • ab177461 (purified) at 1:20 dilution (2µg) immunoprecipitating Pyruvate Dehydrogenase E1-alpha subunit in 293T whole cell lysate.
    Lane 1 (input): 293T (Human embryonic kidney epithelial cell) whole cell lysate 10µg
    Lane 2 (+): ab177461 & 293T whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab177461 in 293T 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat colon tissue sections labeling Pyruvate Dehydrogenase E1-alpha subunit with Purified ab177461 at 1:250 dilution (0.52 µg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/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.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse colon tissue sections labeling Pyruvate Dehydrogenase E1-alpha subunit with Purified ab177461 at 1:250 dilution (0.52 µg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/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.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon tissue sections labeling Pyruvate Dehydrogenase E1-alpha subunit with Purified ab177461 at 1:250 dilution (0.52 µg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/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.
  • All lanes : Anti-Pyruvate Dehydrogenase E1-alpha subunit (phospho S293) antibody [EPR12200] (ab177461) at 1/2000 dilution (purified)

    Lane 1 : Mouse kidney lysates
    Lane 2 : Mouse kidney lysates.Then the membrane was incubated with phosphatase.

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 40 kDa



    Blocking and diluting buffer : 5% NFDM/TBST
  • All lanes : Anti-Pyruvate Dehydrogenase E1-alpha subunit (phospho S293) antibody [EPR12200] (ab177461) at 1/2000 dilution (purified)

    Lane 1 : HT-29 (Human colorectal adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : HT-29 (Human colorectal adenocarcinoma epithelial cell) treated with 8 mM Sodium butyrate for 24 hours whole cell lysates
    Lane 3 : HT-29 (Human colorectal adenocarcinoma epithelial cell) treated with 8 mM Sodium butyrate for 24 hours whole cell lysates.Then the membrane was incubated with phosphatase

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 40 kDa



    Blocking and diluting buffer : 5% NFDM/TBST
  • Serially diluted unpurified ab177461 was bound to immobilised Phospho peptide (S293) - or Control peptide (1 microgram x mL-1). The antibody was detected by HRP-labelled goat anti-rabbit IgG (ab97080; diluted 50000 times) and signal was developed with TMB substrate.

  • All lanes : Anti-Pyruvate Dehydrogenase E1-alpha subunit (phospho S293) antibody [EPR12200] (ab177461) at 1/1000 dilution (unpurified)

    All lanes : HeLa Whole Cell Lysate + Calyculin A (30nM for 20min)

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : goat anti-rabbit (green) and goat anti-mouse at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 40 kDa



    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour and then treated with either buffer (lane 1) or alkaline phosphatase (lane 2), before being incubated with ab177461 overnight at 4°C. Antibody binding was detected using IR-labelled goat anti-rabbit (green) and goat anti-mouse (Red) at 1:10,000 dilution for one hour at room temperature before imaging.

     

  • All lanes : Anti-Pyruvate Dehydrogenase E1-alpha subunit (phospho S293) antibody [EPR12200] (ab177461) at 1/1000 dilution (unpurified)

    Lane 1 : Immunoprecipitation pellet from 293T cells lysate at 10 µg
    Lane 2 : 1X PBS (negative control)

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

    Developed using the ECL technique.

    Predicted band size: 40 kDa

  • Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Pyruvate dehydrogenase E1-alpha subunit with unpurified  ab177461 at 1/50 dilution.

  • Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling Pyruvate dehydrogenase E1-alpha subunit with unpurified  ab177461 at 1/50 dilution.

  • All lanes : Anti-Pyruvate Dehydrogenase E1-alpha subunit (phospho S293) antibody [EPR12200] (ab177461) at 1/1000 dilution (unpurified)

    Lane 1 : 293T cell lysates untreated
    Lane 2 : 293T cell lysates, membrane treated with Lambda Phosphatase.

    Lysates/proteins at 10 µg per lane.

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

    Developed using the ECL technique.

    Predicted band size: 40 kDa

References

This product has been referenced in:
  • Ryan ZC  et al. 1a,25-dihydroxyvitamin D3mitigates cancer cell mediated mitochondrial dysfunction in human skeletal muscle cells. Biochem Biophys Res Commun 496:746-752 (2018). WB . Read more (PubMed: 29366785) »
  • He Z  et al. MiR-422a regulates cellular metabolism and malignancy by targeting pyruvate dehydrogenase kinase 2 in gastric cancer. Cell Death Dis 9:505 (2018). Read more (PubMed: 29725130) »
See all 11 Publications for this product

Customer reviews and Q&As

1-2 of 2 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
Cow Cell lysate - whole cell (VVEC)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
20 µg
Treatment
ATP 100 uM 0,5, 30 min
Specification
VVEC
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jun 03 2016

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