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

概述

  • 产品名称Phospho S232 PDH E1 alpha protein (PDHA1) Profiling ELISA试剂盒
    参阅全部 Pyruvate Dehydrogenase E1-alpha subunit 试剂盒
  • 检测方法Colorimetric
  • 精确度
    批次内
    样品 n Mean SD CV%
    1 8 = 8.3%
    批次间
    样品 n Mean SD CV%
    1 4 = 7.7%
  • 测试
    1 x 96 test
  • 样品类型
    Cell culture extracts, Tissue Extracts
  • 检测类型Sandwich (quantitative)
  • 灵敏度
    = 31 µg/ml
  • 范围
    31 µg/ml - 1000 µg/ml
  • 实验步骤Multiple steps standard assay
  • 种属反应性
    与反应: Mouse, Rat, Cow, Human
  • 产品概述

    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.

  • 说明

    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.

  • 经测试应用适用于: ELISAmore details
  • 平台Microplate

性能

  • 功能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).
  • 组织特异性Ubiquitous.
  • 疾病相关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.
  • 翻译后修饰Phosphorylation at Ser-293 by PDK family kinases blocks the access to active site, and inactivates the enzyme.
  • 细胞定位Mitochondrion matrix.
  • Information by UniProt
  • 别名
    • 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
  • 数据库链接

应用

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.

应用 Ab评论 说明
ELISA Use at an assay dependent concentration.

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

  • 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.

实验方案

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

ab115343 has not yet been referenced specifically in any publications.

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