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AB115345

Phospho S300 PDH E1 alpha protein (PDHA1) Profiling ELISA Kit

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(1 Publication)

Phospho S300 PDH E1 alpha protein (PDHA1) Profiling ELISA Kit is a Sandwich ELISA for the measurement of Phospho S300 PDH E1 alpha protein (PDHA1) Profiling in in Cell/Tissue Extracts samples from a wide range of species.
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Functional Studies - Phospho S300 PDH E1 alpha protein (PDHA1) Profiling ELISA Kit (AB115345)
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Supplier Data

Functional Studies - Phospho S300 PDH E1 alpha protein (PDHA1) Profiling ELISA Kit (AB115345)

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 did not show a significant endogenous phosphorylation signal at S300 this could be increased by kinase treatment. Phosphatase treatement had no effect on the endogenous (low) level of phospho S300 signal.

Functional Studies - Phospho S300 PDH E1 alpha protein (PDHA1) Profiling ELISA Kit (AB115345)
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Supplier Data

Functional Studies - Phospho S300 PDH E1 alpha protein (PDHA1) Profiling ELISA Kit (AB115345)

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 did not reduce the level of phospho S300 confirming that there is little endogenous phospho S300. Conversely NaF treatment, to inhibit cellular serine phosphatases, did increase the phosphorylation level of S300.

ELISA - Phospho S300 PDH E1 alpha protein (PDHA1) Profiling ELISA Kit (AB115345)
  • ELISA

Supplier Data

ELISA - Phospho S300 PDH E1 alpha protein (PDHA1) Profiling ELISA Kit (AB115345)

The phosphorylation state of PDHA1 can vary by treatment but also by cell culture conditions such as media supplements, nutrients and also cell density.

Key facts

Detection method

Colorimetric

Sample types

Cell culture extracts, Tissue Extracts

Reacts with

Mouse, Rat, Cow, Human

Assay type

Sandwich

Results type

Quantitative

Sensitivity

= 15 µg/mL

Range

15 - 500 µg/mL

Assay Platform

Microplate

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

ab115345 is an in vitro enzyme-linked immunosorbent assay to determine the levels of phospho S300 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 S300 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 S300 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.

Store all components at 4°C. This kit is stable for 6 months from

receipt. Unused microplate strips should be returned to the pouch

containing the desiccant and resealed.

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Precision

[ { "reproducibilityType": "Inter", "sample": "Sample 1", "replicates": 4, "mean": null, "standardDeviation": null, "coefficientOfVariability": "7.8" }, { "reproducibilityType": "Intra", "sample": "Sample 1", "replicates": 8, "mean": null, "standardDeviation": null, "coefficientOfVariability": "2.5" } ]
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What's included?

{ "values": { "1x96Tests": { "sellingSize": "1 x 96 Tests", "publicAssetCode":"ab115345-1x96Tests", "assetComponentDetails": [ { "size":"1 x 6 mL", "name":"10X Blocking Buffer", "number":"AB115345-CMP06", "productcode":"" }, { "size":"1 x 0.7 mL", "name":"10X phospho S300 PDHA1 protein Detector Antibody", "number":"AB115345-CMP03", "productcode":"" }, { "size":"1 x 20 mL", "name":"20X Buffer", "number":"AB115345-CMP04", "productcode":"" }, { "size":"1 x 12 mL", "name":"HRP Development Solution", "number":"AB115345-CMP01", "productcode":"" }, { "size":"1 x 1 mL", "name":"10X HRP Label", "number":"AB115345-CMP02", "productcode":"" }, { "size":"1 x 15 mL", "name":"Extraction Buffer (ab260490)", "number":"AB115345-CMP05", "productcode":"" }, { "size":"1 x 1 Unit", "name":"Microplate 96 antibody coated wells in 12 strips", "number":"AB115345-CMP07", "productcode":"" } ] } } }
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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

PDHA1 also known as the pyruvate dehydrogenase E1 alpha subunit plays a mechanical role in cellular metabolism. It forms part of the larger pyruvate dehydrogenase (PDH) complex where it serves as a critical catalytic component. PDHA1 is expressed ubiquitously across different tissue types reflecting its fundamental function in energy production. The molecular weight of the PDHA1 protein is approximately 43 kDa. Alternate names for this protein include the PDH E1 component and it partners closely with other components in the PDH complex to facilitate its role.
Biological function summary

PDHA1 engages in the conversion of pyruvate into acetyl-CoA an important step in cellular respiration. This protein is part of the PDH complex which consists of multiple copies of three catalytic and two regulatory subunits. The conversion process is essential for linking glycolysis to the citric acid cycle efficiently channeling energy substrates within the cell. Furthermore the functional activity of PDHA1 is regulated through phosphorylation by the pyruvate dehydrogenase kinases (PDKs) and dephosphorylation by PDH phosphatases.

Pathways

PDHA1 is integral to the metabolic pathway of cellular respiration and energy production. It enables the transition between glycolysis and the citric acid cycle by facilitating the conversion of pyruvate to acetyl-CoA which enters the citric acid cycle. Related proteins in this pathway include PDHA2 and the regulatory PDKs that modulate PDHA1 activity. These interactions ensure energy metabolism adapts to various cellular conditions influencing energy balance and substrate utilization.

Mutations or dysfunctions in PDHA1 can lead to disorders such as pyruvate dehydrogenase deficiency and Leigh syndrome. These conditions result from impaired energy metabolism leading to severe neurological symptoms and overall energy deficits in tissues with high metabolic demands. The link between PDHA1 and diseases highlights the importance of maintaining its function. Additionally altered interaction with proteins involved in phosphorylation such as the PDKs can exacerbate pathogenic conditions by further disbalancing metabolic activities.
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Product protocols

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Target data

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Publications (1)

Recent publications for all applications. Explore the full list and refine your search

Molecular systems biology 13:953 PubMed29109155

2017

Temporal fluxomics reveals oscillations in TCA cycle flux throughout the mammalian cell cycle.

Applications

Unspecified application

Species

Unspecified reactive species

Eunyong Ahn,Praveen Kumar,Dzmitry Mukha,Amit Tzur,Tomer Shlomi
View all publications
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websiteProtocolBooklet
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