The Tricarboxylic Acid Cycle Sampler Kit provides an economical means of detecting select components involved in tricarboxylic acid cycle. The kit contains enough primary antibodies to perform at least two western blot experiments per antibody.
Specificity / Sensitivity
Each antibody in this kit recognizes endogenous levels of its specific target protein. IDH1 (D2H1) Rabbit mAb does not recognize endogenous IDH2 protein, but does recognize IDH2 when recombinantly overexpressed. IDH2 (D8E3B) Rabbit mAb does not cross-react with IDH1 protein. MPC1 (D2L9I) Rabbit mAb does not cross-react with MPC2 protein. MPC2 (D4I7G) Rabbit mAb does not cross-react with MPC1 protein.
Source / Purification
Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly540 of human ACO2 protein, residues near the carboxy terminus of human citrate synthase protein, residues surrounding Pro188 of human DLST protein, residues surrounding Gly354 of human fumarase protein, residues surrounding Arg222 of human IDH1 protein, residues surrounding Val195 of human IDH2 protein, residues near the carboxy terminus of human MPC1 protein, residues surrounding Asn33 of human MPC2 protein and residues surrounding Gly166 of human SDHA protein, respectively.
The tricarboxylic acid (TCA) cycle includes various enzymatic reactions that constitute a key part of cellular aerobic respiration. The transport of the glycolytic end product pyruvate into mitochondria and the decarboxylation of pyruvate in the TCA cycle generate energy through oxidative phosphorylation under aerobic conditions (1,2). Two inner mitochondrial membrane proteins, mitochondrial pyruvate carrier 1 (MPC1) and mitochondrial pyruvate carrier 2 (MPC2), form a 150 kDa complex and are essential proteins in the facilitated transport of pyruvate into mitochondria (1,2). Citrate synthase catalyzes the first and rate-limiting reaction of the TCA cycle (3). Mitochondrial aconitase 2 (ACO2) catalyzes the conversion of citrate to isocitrate via cis-aconitate (4). IDH1 and IDH2 are two of the three isocitrate dehydrogenases that catalyze oxidative decarboxylation of isocitrate to α-ketoglutarate (α-KG) (5). IDH1 functions as a tumor suppressor in the cytoplasm and peroxisomes, whereas IDH2 is in mitochondria and is involved in the TCA cycle (5). Mutations in IDH2 have also been identified in malignant gliomas (6). Dihydrolipoamide succinyltransferase (DLST) is a subunit of the α-ketoglutarate dehydrogenase complex, a key enzymatic complex in the TCA cycle (7). Succinate dehydrogenase subunit A (SDHA) is a component of the TCA cycle and the electron transport chain and is involved in the oxidation of succinate (8). Fumarase catalyzes the conversion of fumarate to malate (9). Fumarase deficiency leads to the accumulation of fumarate, an oncometabolite that has been shown to promote epithelial-to-mesenchymal-transition (EMT), a developmental process that has been implicated in oncogenesis (10).