PDK2 ELISA Kit (Mouse)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Pyruvate Dehydrogenase Kinase Isozyme 2 (PDK2) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Pyruvate Dehydrogenase Kinase Isozyme 2 (PDK2) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

[Pyruvate dehydrogenase (acetyl-transferring) ] kinase isozyme 2, mitochondrial; [Pyruvate dehydrogenase [lipoamide]] kinase isozyme 2, mitochondrial; PDH kinase 2; pyruvate dehydrogenase 2; Pyruvate dehydrogenase kinase isoform 2; pyruvate dehydrogenase kinase, isoenzyme 2; pyruvate dehydrogenase kinase, isozyme 2.

Kinase that plays a key role in the regulation of glucose and fatty acid metabolism and homeostasis via phosphorylation of the pyruvate dehydrogenase subunits PDHA1 and PDHA2. This inhibits pyruvate dehydrogenase activity, and thereby regulates metabolite flux through the tricarboxylic acid cycle, down-regulates aerobic respiration and inhibits the formation of acetyl-coenzyme A from pyruvate. Inhibition of pyruvate dehydrogenase decreases glucose utilization and increases fat metabolism. Mediates cellular responses to insulin. Plays an important role in maintaining normal blood glucose levels and in metabolic adaptation to nutrient availability. Via its regulation of pyruvate dehydrogenase activity, plays an important role in maintaining normal blood pH and in preventing the accumulation of ketone bodies under starvation. Plays a role in the regulation of cell proliferation and in resistance to apoptosis under oxidative stress. Plays a role in p53/TP53-mediated apoptosis.

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Pyruvate Dehydrogenase Kinase Isozyme 2 (PDK2) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Pyruvate Dehydrogenase Kinase Isozyme 2 (PDK2) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Pyruvate Dehydrogenase Kinase Isozyme 2 (PDK2), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Pyruvate Dehydrogenase Kinase Isozyme 2 (PDK2) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Matrix Recovery Range (%) Average (%) serum (n=5) 83-97 88 EDTA plasma (n=5) 84-99 87 heparin plasma (n=5) 96-105 101 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) serum (n=5) 83-97 88 serum (n=5) 83-97 88 EDTA plasma (n=5) 84-99 87 EDTA plasma (n=5) 84-99 87 heparin plasma (n=5) 96-105 101 heparin plasma (n=5) 96-105 101

Component Amount Anti-Pdk2 Microplate 96 Wells (12 x 8 Well Strips) Pdk2 Lyophilized Standard 2 x 10 ng 100X Biotinylated Pdk2 Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Component Amount Component Amount Anti-Pdk2 Microplate 96 Wells (12 x 8 Well Strips) Anti-Pdk2 Microplate 96 Wells (12 x 8 Well Strips) Pdk2 Lyophilized Standard 2 x 10 ng Pdk2 Lyophilized Standard 2 x 10 ng 100X Biotinylated Pdk2 Detector Antibody 120 uL 100X Biotinylated Pdk2 Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL

This assay has high sensitivity and excellent specificity for detection of Pyruvate Dehydrogenase Kinase Isozyme 2 (PDK2) . No significant cross-reactivity or interference between Pyruvate Dehydrogenase Kinase Isozyme 2 (PDK2) and analogues was observed.