Phospho S232 PDH E1 alpha protein (PDHA1) Profiling ELISA Kit (ab115343)

製品の概要

  • 製品名
    Phospho S232 PDH E1 alpha protein (PDHA1) Profiling ELISA Kit
    Pyruvate Dehydrogenase E1-alpha subunit キット 製品一覧
  • 検出方法
    Colorimetric
  • 再現性
    Intra-Assay(同時再現性)
    サンプル N 平均値 SD CV%
    1 8 = 8.3%
    Inter-Assay(日差再現性)
    サンプル N 平均値 SD CV%
    1 4 = 7.7%
  • サンプルの種類
    Cell culture extracts, Tissue Extracts
  • アッセイタイプ
    Sandwich (quantitative)
  • 検出感度
    = 31 µg/ml
  • 検出範囲
    31 µg/ml - 1000 µg/ml
  • ステップ
    Multiple steps standard assay
  • 種交差性
    交差種: Mouse, Rat, Cow, Human
  • 製品の概要

    ab115343 is an in vitro enzyme-linked immunosorbent assay to determine the levels of phospho S232 PDHA1 protein in cell and tissue lysates. The assay employs a mouse antibody specific for PDHA1 protein coated on a 96-well plate. Samples are pipetted into the wells and PDHA1 protein present in the sample is bound to the wells by the immobilized antibody. The wells are washed and a rabbit anti-phospho S232 PDHA1 protein detector antibody is added. After washing away unbound detector antibody, HRP-conjugated anti-rabbit antibody is pipetted into the wells. The wells are again washed, an HRP substrate solution (TMB) is added to the wells and color develops in proportion to the amount of phospho S232 PDHA1 protein bound. The developing blue color is measured at 600 nm. Optionally the reaction can be stopped by adding hydrochloric acid which changes the color from blue to yellow and the intensity can be measured at 450 nm.

  • 特記事項

    Store all components at 4°C. The kits are stable for at least 6

    months. Unused microplate strips should be returned to the pouch

    containing the desiccant and resealed.

  • アプリケーション
    適用あり: ELISAmore details
  • 試験プラットフォーム
    Microplate

製品の特性

  • 保存方法
    Store at +4°C. Please refer to protocols.
  • 内容 1 x 96 tests
    10X Blocking Buffer 1 x 6ml
    10X HRP Label 1 x 1ml
    10X phospho S232 PDHA1 protein Detector Antibody 1 x 0.7ml
    20X Buffer 1 x 20ml
    Extraction Buffer 1 x 15ml
    Microplate 96 antibody coated wells in 12 strips 1 unit
    HRP Development Solution 1 x 12ml
  • 研究分野
  • 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 dehydrogenase E1-alpha deficiency (PDHAD) [MIM:312170]. An enzymatic defect causing primary lactic acidosis in children. It is associated with a broad clinical spectrum ranging from fatal lactic acidosis in the newborn to chronic neurologic dysfunction with structural abnormalities in the central nervous system without systemic acidosis.
    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.
  • 翻訳後修飾
    Phosphorylation at Ser-293 by PDK family kinases blocks the access to active site, and inactivates the enzyme.
  • 細胞内局在
    Mitochondrion matrix.
  • Information by UniProt
  • 別名
    • mitochondrial
    • ODPA_HUMAN
    • PDHA1
    • PDHCE1A
    • PDHE1-A type I
    • PHE1A
    • Pyruvate dehydrogenase (lipoamide) alpha 1
    • Pyruvate dehydrogenase complex E1 alpha polypeptide 1
    • Pyruvate dehydrogenase E1 component subunit alpha
    • Pyruvate dehydrogenase E1 component subunit alpha somatic form mitochondrial
    • Pyruvate dehydrogenase, alpha-1
    • somatic form
    see all
  • 参照データベース

アプリケーション

Our Abpromise guarantee covers the use of ab115343 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
ELISA Use at an assay dependent concentration.

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  • The PDHA1 bound from undosed HeLa cells was subject to in-well kinase treatment (PDK1&3) or in-well phosphatase treatment (PDP1) according to the supplementary protocol shown below. Untreated cells showed a significant endogenous phosphorylation signal at S232 which is only slightly increased by kinase treatment. Conversely phosphatase treatment was able to significantly reduce the phospho S232 signal from the endogenous levels.

  • HeLa cells were cultured for 4 hours in media supplemented with DCA (20mM) to specifically inhibit mitochondrial PDH kinases, or NaF (20mM), a general inhibitor of serine/threonine protein phosphatases. The DCA treatment reduced the level of phospho S232. Conversely NaF treatment, to inhibit cellular serine phosphatases, increased the phosphorylation level of S232.
  • Example control sample curve for HepG2 cell extract. The phosphorylation state of PDHA1 can vary by treatment but also by cell culture conditions such as media supplements, nutrients and also cell density.

  • Example control sample curve for HeLa cell extract. The phosphorylation state of PDHA1 can vary by treatment but also by cell culture conditions such as media supplements, nutrients and also cell density.

プロトコール

参考文献

ab115343 has not yet been referenced specifically in any publications.

レビューと Q&A

Unfortunately we are not able to sell the detection antibodies from this kit individually.

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

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