AAMT008 Sigma-AldrichPDH Activity Assay Kit

This product has been discontinued.

Product Description

The PDH Activity Assay Kit is used to determine the activity of pyruvate dehydrogenase (PDH) in a sample. The PDH enzyme is immunocaptured within the wells of the microplate and PDH activity is determined by following the reduction of NAD⁺ to NADH, coupled to the reduction of a reporter dye to yield a colored (yellow) reaction product whose concentration can be monitored by measuring the increase in absorbance at 450 nm (See Figure 1 at the bottom of this page).

PDH is the key regulatory enzyme of cellular metabolism because it links the TCA cycle and subsequent oxidative phosphorylation with glycolysis and gluconeogenesis as well as with both lipid and amino acid metabolism. PDH activity is regulated by PDK-dependent phosphorylation and PDP dependent dephosphorylation of PDH. Phosphorylation inactivates PDH whereas dephosphorylation activates PDH.

The PDH Activity Assay Kit can be used to elucidate various aspects of PDH activity, physiologic regulation and phosphorylation status. Please read TB561 for three possible applications for this kit.

• 15 ml         20X Buffer, (72545-15ML)
• 2 x 1 ml     Detergent, (72546-1ML)
• 2 x 600 µl  20X Reagent Mix, (72547-600UL)
• 1 ea           Microplate, (72548-1EA)
• 13 ml        5X Stabilizer, (72549-1EA)
• 250 µl       100X Coupler, (72550-250UL)
• 250 µl       100X Reagent Dye, (72551-250UL)

*This kit contains sufficient materials for 96 measurements (one 96-well microplate).

Storage: Store all components at 4°C. Store the 100X Coupler at –80°C. This kit is stable for at least 6 months.

This kit contains the necessary buffers and reagents for the respective reaction. The kit also contains a 96-well microplate with a monoclonal antibody pre-bound to the wells of the microplate. This plate can be broken into 12 separate 8-well strips for convenience; therefore the plate can be used for up to 12 separate experiments.