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Pdha1

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PDHA1 is a protein that catalyzes the conversion of pyruvate to acetyl-CoA, a key step in cellular respiration. It is an essential component of the pyruvate dehydrogenase complex.

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Lab products found in correlation

Pdhk4, by Novus Biologicals (2 mentions) Pdhk2, by Novus Biologicals (2 mentions) Pdhk3, by Novus Biologicals (2 mentions) Gapdh, by Abcam (2 mentions) Hif 1α, by BD (2 mentions) Sirt5, by Cell Signaling Technology (1 mentions) Succinyl lysine antibody, by PTM Biolabs (1 mentions) Fluorescently conjugated secondary antibodies, by LI COR (1 mentions) Tbs blocking buffer, by LI COR (1 mentions)

Close spelling variants of this product

P-PDHA1 PDHA1 antibody

6 protocols using pdha1

1

Detecting PDHA1 Expression in Cancers

Cited 1 time
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Of these Riskgenes, PDHA1 plays a crucial role in multiple malignancies as a tumor suppressor or oncogene. We detected the expression pattern of PDHA1 (1:500, Abcam) in human clinical samples using immunofluorescence, which was carried out as we described previously (34 (link)).
Tian X.M., Xiang B., Yu Y.H., Li Q., Zhang Z.X., Zhanghuang C., Jin L.M., Wang J.K., Mi T., Chen M.L., Liu F, & Wei G.H. (2022). A novel cuproptosis-related subtypes and gene signature associates with immunophenotype and predicts prognosis accurately in neuroblastoma. Frontiers in Immunology, 13, 999849.
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2

Porcine Heart PDC Desuccinylation Assay

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Porcine heart PDC (Sigma Aldrich, St. Louis, MO) was purified by centrifugation at 135,000×g for 2 hours in 100 mM potassium phosphate, pH 7.5, 0.05% lauryl maltoside, 2.5 mM EDTA, and 30% glycerol. Desuccinylation reactions were carried out on 30 µg of purified porcine heart PDC in a final reaction volume of 60 µl in presence of 25 mM Tris-Cl, pH 8.0, 200 mM NaCl, 5 mM KCl, 1 mM MgCl2, 0.1% PEG 8000, and 3.125 mM NAD+ at 37°C for 2 hours. Where indicated, 10 µg of SIRT5 or SIRT5H158Y (expressed and purified in house) was added. During incubation, tubes were occasionally agitated. Following desuccinylation, 15 µl each reaction was analyzed by immunoblotting with a succinyl-lysine antibody (PTM Biolabs, Inc, Chicago, IL). After analysis, the membrane was stripped and re-probed for PDHA1 (Abcam, Cambridge, MA) and SIRT5 (Cell Signaling Technology, Danvers, MA).
Guetschow E.D., Kumar S., Lombard D.B, & Kennedy R.T. (2015). Identification of Sirtuin 5 Inhibitors by Ultrafast Microchip Electrophoresis Using Nanoliter Volume Samples. Analytical and bioanalytical chemistry, 408(3), 721-731.
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3

Kinase Activity of PDK1 in Pyruvate Dehydrogenase Regulation

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Kinase activity of PDK was slightly modified as previously described (19 (link),20 (link)). Briefly, 50 ng of recombinant PDK1 (Abcam) was incubated with 100 ng of recombinant pyruvate dehydrogenase E1 subunit alpha 1 (PDHA1; Abcam) for 30 min at 37°C in PDK1 buffer containing 20 mM Tris buffer (pH 7.5), 0.1 mM EDTA, 1 mM MgCl2, 2 mM dithiothreitol (DTT) and 250 µM ATP. The samples were subjected to SDS-PAGE and immunoblotted using antibodies against PDK1, p-PDHA1, and PDHA1 (Table I).
Yang E.S., Do Y., Cheon S.Y., Kim B., Ling J., Cho M.K., Kim T., Bae S.J, & Ha K.T. (2023). Andrographolide suppresses aerobic glycolysis and induces apoptotic cell death by inhibiting pyruvate dehydrogenase kinase 1 expression. Oncology Reports, 49(4), 72.
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4

Western Blot Analysis of Metabolic Enzymes

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Whole cell extracts were fractionated by SDS-PAGE and transferred to
nitrocellulose membrane by wet transfer for 4 h on ice at 60 V. Blots were
blocked in TBS blocking buffer (Li-Cor) and then stained overnight with
primary antibody at 4°C. The next day, membranes were washed three
times in TBST (TBS and 0.1% Tween-20), stained with fluorescently conjugated
secondary antibodies (Li-Cor) at 1:10,000 dilution in TBS blocking buffer
for 1 h at room temperature, and then imaged in the 680- and 800-nm
channels. Antibodies used: Gapdh (CST), Gpi1 (Thermo Fisher), Ldha (CST),
Pdha1 (Abcam), Tpi1 (Abcam), G6pdx (Abcam), a-tubulin (Santa Cruz).
Wu L., Hollinshead K.E., Hao Y., Au C., Kroehling L., Ng C., Lin W.Y., Li D., Silva H.M., Shin J., Lafaille J.J., Possemato R., Pacold M.E., Papagiannakopoulos T., Kimmelman A.C., Satija R, & Littman D.R. (2020). Niche-Selective Inhibition of Pathogenic Th17 Cells by Targeting Metabolic Redundancy. Cell, 182(3), 641-654.e20.
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5

PDH Regulatory Protein Analysis

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Cells were lysed with RIPA buffer containing protease and phosphatase inhibitors (5 mM sodium fluoride, 2 mM β -glycerophosphate, 1 mM sodium orthovanadate, 1 mM phenylmethylsulfonyl fluoride (PMSF), and complete mini protease inhibitor cocktail (Roche)), 20–50 μg of total proteins electrophoresed and blotted to PVDF, and probed with anti: PDHA1 (1:6000, MitoSciences), pSer293-E1α (1:1000, EMD Chemicals), pSer300-E1α (1:1000, EMD Chemicals), pSer232-E1α (1:3000, EMD Chemicals), PDHK1 (1:4000, Assay Designs), PDHK2 (1:500, Novus), PDHK3 (1:1000, Novus), PDHK4 (1:1000, Novus), PDP1 (1:1000, Sigma Aldrich), HIF1α (1:1500, BD), and GAPDH (abcam). Primary antibodies were detected with fluorochrome-labeled secondary antibodies (Li-Cor) visualized on a Li-Cor Odyssey.
Echeverri Ruiz N.P., Mohan V., Wu J., Scott S., Kreamer M., Benej M., Golias T., Papandreou I, & Denko N.C. (2021). Dynamic regulation of mitochondrial pyruvate metabolism is necessary for orthotopic pancreatic tumor growth. Cancer & Metabolism, 9, 39.
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6

Characterizing PDH Complex Regulation

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Cells were lysed with RIPA buffer containing protease and phosphatase inhibitors (5 mM sodium fluoride, 2 mM β -glycerophosphate, 1 mM sodium orthovanadate, 1 mM phenylmethylsulfonyl fluoride (PMSF), and complete mini protease inhibitor cocktail (Roche)), 20-50 μ g of total proteins electrophoresed and blotted to PVDF, and probed with anti: PDHA1 (1:6000, MitoSciences), pSer293-E1α (1:1000, EMD Chemicals), pSer300-E1α (1:1000, EMD Chemicals), pSer232-E1α (1:3000, EMD Chemicals), PDHK1 (1:4000, Assay Designs), PDHK2 (1:500, Novus), PDHK3 (1:1000, Novus), PDHK4 (1:1000, Novus), PDP1 (1:1000, Sigma Aldrich), HIF1α (1:1500, BD), GAPDH (abcam). Primary antibodies were detected with fluorochrome labelled secondary antibodies (Li-Cor) visualized on a Li-Cor Odyssey.
Ruiz N.P.E., Mohan V., Wu J., Scott S., Kreamer M., Benej M., Papandreou I., & Denko N. (2021). Dynamic regulation of mitochondrial pyruvate metabolism Is necessary for orthotopic pancreatic tumor growth.
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