Anti-Pyruvate Dehydrogenase E1-alpha subunit (phospho S293) 抗体 (ab92696)

製品の概要

  • 製品名Anti-Pyruvate Dehydrogenase E1-alpha subunit (phospho S293) antibody
    Pyruvate Dehydrogenase E1-alpha subunit 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to Pyruvate Dehydrogenase E1-alpha subunit (phospho S293)
  • アプリケーション適用あり: WB, ICC/IF, IHC-Pmore details
  • 種交差性
    交差種: Mouse, Human
    交差が予測される動物種: Rat, Cow, Pig, Orangutan
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 250 - 350 of Human Pyruvate Dehydrogenase E1-alpha subunit, phosphorylated at S293.

  • ポジティブ・コントロール
    • This antibody gave a positive signal in Human liver tissue lysate as well as the following whole cell lysates: HeLa; HEK293; HepG2. This antibody gave a positive result in IHC in the following FFPE tissue: Human lung adenocarcinoma.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab92696 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
WB Use a concentration of 1 µg/ml. Detects a band of approximately 43 kDa (predicted molecular weight: 43 kDa).
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 1 µg/ml.

ターゲット情報

  • 機能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).
  • 組織特異性Ubiquitous.
  • 関連疾患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.
  • 細胞内局在Mitochondrion matrix.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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

Anti-Pyruvate Dehydrogenase E1-alpha subunit (phospho S293) antibody 画像

  • Serially diluted ab92696 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

  • All lanes : Anti-Pyruvate Dehydrogenase E1-alpha subunit (phospho S293) antibody (ab92696) at 1 µg/ml

    Lane 1 : HeLa Whole Cell Lysate + Calyculin A (30 nM for 20 min)
    Lane 2 : HeLa Whole Cell Lysate + Calyculin A (30 nM for 20 min)

    Lysates/proteins at 20 µg per lane.

    Secondary
    goat anti-rabbit (green) and goat anti-mouse (red) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 43 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 ab92696 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.

     

  • ab92696 staining Pyruvate Dehydrogenase E1-alpha subunit (phospho S293) in Human HUVEC cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 pH 7.4 for 5 minutes and blocked with 5% BSA for 20 minutes at room temperature. Samples were incubated with primary antibody (1/500 in PBS) for 1 hour . A CF488-conjugated Donkey anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.

    See Abreview

  • All lanes : Anti-Pyruvate Dehydrogenase E1-alpha subunit (phospho S293) antibody (ab92696) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 4 : Human liver tissue lysate - total protein (ab29889)
    Lane 5 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Modified blocking peptide at 1 µg/ml
    Lane 6 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate with Modified blocking peptide at 1 µg/ml
    Lane 7 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate with Modified blocking peptide at 1 µg/ml
    Lane 8 : Human liver tissue lysate - total protein (ab29889) with Modified blocking peptide at 1 µg/ml
    Lane 9 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Non-modified blocking peptide at 1 µg/ml
    Lane 10 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate with Non-modified blocking peptide at 1 µg/ml
    Lane 11 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate with Non-modified blocking peptide at 1 µg/ml
    Lane 12 : Human liver tissue lysate - total protein (ab29889) with Non-modified blocking peptide at 1 µg/ml

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 43 kDa
    Observed band size : 43 kDa
    Additional bands at : 34 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 2 minutes
  • ICC/IF image of ab92696 stained HepG2 cells. The cells were 100% methanol fixed (5 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 (ab92696, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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. This antibody also gave a positive result in 100% methanol fixed (5 min) HeLa cells at 1µg/ml.

  • IHC image of Pyruvate Dehydrogenase E1-alpha subunit (phospho S293) staining in Human lung adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab92696, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

Anti-Pyruvate Dehydrogenase E1-alpha subunit (phospho S293) antibody (ab92696) 使用論文

This product has been referenced in:
  • Wang CW  et al. In vivo genetic dissection of tumor growth and the Warburg effect. Elife 5:N/A (2016). Read more (PubMed: 27585295) »
  • Qin XY  et al. The effect of acyclic retinoid on the metabolomic profiles of hepatocytes and hepatocellular carcinoma cells. PLoS One 8:e82860 (2013). WB ; Human . Read more (PubMed: 24376596) »

See all 3 Publications for this product

Product Wall

Application Western blot
Sample Human Cell lysate - whole cell (Vascular Smooth Muscle Cell)
Gel Running Conditions Reduced Denaturing
Loading amount 25 µg
Specification Vascular Smooth Muscle Cell
Blocking step Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5µg/mL · Temperature: 21°C
Username

Abcam user community

Verified customer

投稿 Jul 20 2016

Application Western blot
Sample Sheep Tissue lysate - whole (liver)
Gel Running Conditions Reduced Denaturing
Loading amount 30 µg
Specification liver
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Jennifer Bruce

Verified customer

投稿 Jun 15 2015

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (10%)
Sample Human Cell lysate - whole cell (Human lung adenocarcinoma epithelial cell line)
Specification Human lung adenocarcinoma epithelial cell line
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Stamatia Pouliliou

Verified customer

投稿 Mar 23 2015

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 20 minute(s) · Concentration: 5%
Sample Human Cell (HuVEC)
Specification HuVEC
Permeabilization Yes - 0.1% v/v Triton X-100 pH 7.4 for 5 min at RT
Fixative Paraformaldehyde
Username

Dr. Dimitra Kalamida

Verified customer

投稿 Nov 18 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Tissue lysate - whole (Heart, left ventricle)
Loading amount 40 µg
Specification Heart, left ventricle
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Dawn Bowles

Verified customer

投稿 Dec 27 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - other (HeLa and U2OS: Purified mitochondria lysate from 9)
Loading amount 3e+006 cells
Specification HeLa and U2OS: Purified mitochondria lysate from 9
Gel Running Conditions Reduced Denaturing (8% polyacrylamide gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

投稿 Aug 09 2012

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"