title: Immunocapture and microplate-based activity and quantity measurement of pyruvate dehydrogenase in human peripheral blood mononuclear cells.
abstract: BACKGROUND
  Pyruvate dehydrogenase (PDH) activity is altered in many human disorders. Current methods require tissue samples and yield inconsistent results. We describe a modified method for measuring PDH activity from isolated human peripheral blood mononuclear cells (PBMCs). RESULTS/METHODOLOGY: We found that PDH activity and quantity can be successfully measured in human PBMCs. Freeze-thaw cycles cannot efficiently disrupt the mitochondrial membrane. Processing time of up to 20 h does not affect PDH activity with proteinase inhibitor addition and a detergent concentration of 3.3% showed maximum yield. Sample protein concentration is correlated to PDH activity and quantity in human PBMCs from healthy subjects.
  
  
  CONCLUSIONS
  Measuring PDH activity from PBMCs is a novel, easy and less invasive way to further understand the role of PDH in human disease.
authors.affiliation: Department of Emergency Medicine, Beth Israel Deaconess Medical Center, One Deaconess Road, W/CC 2, Boston, MA 02215, USA.
authors.forename: Xiaowen
authors.initials: X
authors.lastname: Liu
authors.forename: Hira
authors.initials: H
authors.lastname: Pervez
authors.forename: Lars W
authors.initials: LW
authors.lastname: Andersen
authors.forename: Amy
authors.initials: A
authors.lastname: Uber
authors.forename: Sophia
authors.initials: S
authors.lastname: Montissol
authors.forename: Parth
authors.initials: P
authors.lastname: Patel
authors.forename: Michael W
authors.initials: MW
authors.lastname: Donnino
mesh_terms: D006801:Humans; D007963:Leukocytes, Mononuclear / Q000276:immunology*; D011768:Pyruvate Dehydrogenase Complex / Q000276:immunology*
chemical_list: D011768:Pyruvate Dehydrogenase Complex
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