MILLIPLEX MAP Multi-species Pyruvate Dehydrogenase (PDH) Complex Magnetic Bead Panel - Cellular Metabolism Multiplex Assay

MILLIPLEX MAP Multi-species Pyruvate Dehydrogenase (PDH) Complex Magnetic Bead Panel, PDHMAG-13K

MILLIPLEX

96-well plate

MILLIPLEX MAP Multi-species Pyruvate Dehydrogenase (PDH) Complex Magnetic Bead Panel, containing PDH (total), PDH (pS232), PDH (pS293), and PDH (pS300) is available in premix format only, and may be used for the analysis of cell lysate or tissue extract.

The Pyruvate Dehydrogenase Complex (PDH Complex or PDC) is a key regulator of cellular energy metabolism, and has roles in numerous diseases including cancer, neurodegeneration, cardiovascular disease, and diabetes. PDH catalyzes the irreversible conversion of pyruvate to acetyl-CoA. PDH activity influences the shift between glycolytic energy production (more prevalent in cancer) and the Krebs cycle and subsequent oxidative phosphorylation, in response to various stimuli, and thereby reverses the aerobic glycolysis (also known as Warburg effect) that is a feature of many cancers.

PDH Complex activity is regulated by PDKs (PDH kinases) and PDPs (PDH phosphatases) which act on three phosphorylation sites on the E1α subunit: Ser232, Ser293, and Ser300. Phosphorylation is inversely correlated with activity and therefore is a useful readout for monitoring the regulation/activity of the enzyme in response to stimuli such as glucose deprivation or hypoxia, as well as in response to treatment with drugs such as PDK inhibitors. For example, one of the modes of action for dichloroacetate (DCA), a potential treatment for cancer and lactic acidosis, is to inhibit PDK activity resulting in decreased PDH Complex phosphorylation and therefore increased PDH activity.

This MILLIPLEX® MAP Multi-species Pyruvate Dehydrogenase (PDH) Complex Magnetic Bead Panel is a Cellular Metabolism multiplex assay. The analytes available for this multiplex kit are: PDH (total), PDH (pS232), PDH (pS293), PDH (pS300).

Multiplexing

• Two hours incubation is recommended for best results.
• These assays require 25 µL diluted cell lysate.
• These kits must be run using the provided Assay Buffer.
• We recommend using 2.5-15 µg cell lysate/well.

• Human
• Mouse
• Rat

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Recommended storage for kit components is 2 - 8°C.

MILLIPLEX® MAP Multi-species Pyruvate Dehydrogenase (PDH) Complex Panel is specific to human, rat, and mouse.

MILLIPLEX MAP Multi-species Pyruvate Dehydrogenase (PDH) Complex Magnetic Bead Panel, PDHMAG-13K

The phospho- and site-specificity of each phospho-assay: PDH (pS232), PDH (pS293), or PDH (pS300) was determined by peptide competition. Phospho-peptides and the corresponding unphosphorylated peptide controls for each of the 3 phospho-target sequences (pSer232, pSer293, and pSer300) were tested for their ability to inhibit/compete away the signal in the multiplex assay. Each assay was shown to be phospho-specific for the corresponding phosphorylation site on the PDH complex demonstrated by significant inhibition of the signal by the site-specific phospho-peptide, while the signal remained largely unchanged from the competition by the unphosphorylated form of the peptide or the other two site-specific phospho-peptides.

PDH (pS232), PDH (pS293), PDH (pS300), PDH (total)

Premixed

MAGNETIC Cellular Metabolism

• Intra- and Inter-Assay CVs based on MFI values.
• Inter-assay: < 10%
• Intra-assay: < 6%

Luminex xMAP

Everything you need in a single kit.