Pyruvate dehydrogenase (PDH) Protein Quantity Dipstick Assay Kit (ab109883)

Overview

• Product namePyruvate dehydrogenase (PDH) Protein Quantity Dipstick Assay KitSee all Pyruvate dehydrogenase kits ...
• Sample type
  Cell culture extracts, Tissue
• Assay typeQuantitative
• Species reactivity
  Reacts with: Mouse, Rat, Cow, Human
  
• Product overview

  ab109883 (MSP31) is used to quantify the activity of the PDH enzyme complex (pyruvate dehydrogenase) from human, bovine, mouse, and rat samples.

  
  PDH is present in all tissues; it plays a central role in metabolism as it is a key regulatory enzyme that functions at the junction between glycolysis and the tricarboxylic acid cycle. The PDH complex is composed of multiple copies of three catalytic component enzymes; pyruvate dehydrogenase or E1 (EC 1.2.4.1), dihydrolipoamide transacetylase or E2 (EC 2.3.1.12) and dihydrolipoamide dehydrogenase or E3 (EC 1.8.1.4). Inherent to its regulatory function, a number of other proteins regulate PDH activity. One of these, dihydrolipoamide dehydrogenase-binding protein (E3Bp) is necessary for the interaction of the E2 and E3 components.
  

  This dipstick assay kit utilizes two monoclonal antibodies. One recognizes the E2 enzyme (monoclonal antibody bound to the dipstick) and the other antibody (gold-conjugated) recognizes both the E2 and E3Bp components of the PDH complex. The signal intensity is measured by a dipstick reader or analyzed by other imaging systems such as a flatbed scanner. The kit is compatible with a variety of sample types (tissue or cell culture) from a number of different species (human, bovine, mouse, and rat).

• Notes

  All components are stable in their provided containers at room temperature out of direct sunlight.

  After diluting the 10X Blocking Buffer to 2X, store at 4°C.

  For long-term storage, all buffers can be stored at 4°C.

• Tested applicationsSandwich ELISA more details

Properties

• Storage instructionsStore at +4°C. Please refer to protocols.

• Components	90 tests	30 tests
  Buffer B (10X Blocking solution)	1 x 1.2ml	1 x 0.4ml
  Buffer C (Wash buffer)	1 x 3ml	1 x 1ml
  Dipsticks	1 x 90 units	1 x 30 units
  Extraction Buffer	1 x 45ml	1 x 15ml
  Gold-conjugated antibody (dried in microplate wells)	1 x 90 tests	1 x 30 tests

• Research Areas
  • Metabolism
  • Pathways and Processes
  • Metabolic signaling pathways
  • Carbohydrate metabolism

  • Metabolism
  • Pathways and Processes
  • Mitochondrial Metabolism
  • Mitochondrial markers

  • Metabolism
  • Pathways and Processes
  • Metabolic signaling pathways
  • Energy transfer pathways
  • Energy Metabolism

  • Cancer
  • Cancer Metabolism
  • Metabolic signaling pathway
  • Metabolism of carbohydrates

  • Kits/ Lysates/ Other
  • Kits
  • Cell Metabolism Kits
  • Other Metabolism Assay

  • Signal Transduction
  • Metabolism
  • Energy Metabolism

  • Signal Transduction
  • Metabolism
  • Mitochondrial

Applications

Pyruvate dehydrogenase (PDH) Protein Quantity Dipstick Assay Kit images

• 

  ELISA - Pyruvate dehydrogenase (PDH) Protein Quantity Dipstick Assay Kit (ab109883)
  An example using ab109883 to quantify PDH levels using various concentration of rat tissue protein extract. Shown is a 1:2 dilution series using a positive control sample. Approximately 7 to 8 dipsticks are suitable for covering the entire working range and the blank for background levels. In this example the dilution series starts with 20 µg of rat heart tissue extract. A one-site hyperbola line was generated for best-fit analysis using GraphPad.
• 

  ELISA - Pyruvate dehydrogenase (PDH) Protein Quantity Dipstick Assay Kit (ab109883)
  An example using ab109883 to quantify PDH levels using various concentration of rat tissue protein extract. Based on the above standard curve values, 18 µg (control - undiluted), 9 µg (sample A - diluted 1:2) and 1.8 µg (sample B - diluted 1:10) of extract were loaded to each dipstick. Two dipsticks per sample were run (typical intra-assay precision is 90-98%). Using GraphPad software, the signal intensity from the standard curve was interpolated and the quantity of PDH in samples A and B was determined. Based on the above analysis, the unknown samples, A and B, had 48% and 9% of control PDH levels, respectively.
• 

  Sandwich ELISA - Pyruvate dehydrogenase (PDH) Protein Quantity Dipstick Assay Kit (ab109883)
  Dipstick assays use the well-established lateral flow concept, whereby capture antibodies are striped onto nitrocellulose membrane and a Whatman paper wicking pad draws the sample through the antibody bands. Detector antibodies, conjugated to gold, are dried in the wells of a 96-well plate. Sample is added to the well, the dipstick inserted, and within minutes the line for each target is revealed as the protein-detector antibody-gold complex binds with the capture antibodies. Multiplexing dipstick assays have multiple target protein lines. A positive control goat anti-mouse antibody line is included on all assays to ensure that adequate wicking of the sample occurred.

Protocols

Protocol Booklet