Branched-Chain Amino Acid Metabolism Antibody Sampler Kit #34995
Product Information
Kit Usage Information
Protocols
Product Description
The Branched-Chain Amino Acid Metabolism Antibody Sampler Kit provides an economical means of detecting select components involved in the branched-chain amino acid (BCAA) metabolism pathway. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.
Specificity / Sensitivity
Each antibody in the Branched-Chain Amino Acid Metabolism Antibody Sampler Kit detects endogenous levels of its target protein. BCAT1 (D6D4K) Rabbit mAb does not cross-react with BCAT2 protein. BCAT2 (D8K3O) Rabbit mAb does not cross-react with BCAT1 protein. Phospho-BCKDH-E1α (Ser293) (E2V6B) Rabbit mAb recognizes endogenous levels of BCKDH-E1α protein only when phosphorylated at Ser292 of mature human BCKDH-E1α, Ser293 of mature mouse BCKDH-E1α, or Ser293 of mature rat BCKDH-E1α.
Source / Purification
Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues near the amino terminus of human BCAT1 protein, residues surrounding Leu39 of human BCAT2 protein, residues near the carboxy terminus of human BCKDH-E1α protein, and a synthetic phosphopeptide corresponding to residues surrounding Ser292 of mature human BCKDH-E1α protein, which corresponds to Ser293 of mature mouse BCKDH-E1α protein and Ser293 of mature rat BCKDH-E1α protein.
Background
BCAT1 and BCAT2 are cytosolic and mitochondrial branched-chain aminotransferases, respectively (1,2). Research studies have implicated BCAT1 in distant metastasis in patients with advanced colorectal cancer (3). Disruption of BCAT2 in mice leads to higher levels of plasma branched-chain amino acids (BCAAs), reduced adiposity and body weight, and increased energy expenditure, suggesting its role in regulating insulin sensitivity (4). BCAAs leucine, isoleucine, and valine are essential amino acids in mammals, but elevated levels of BCAAs have been implicated in cardiovascular and metabolic disorders (5). The branched-chain α-keto acid dehydrogenase complex (BCKDH) catalyzes the rate-limiting step in the BCAA degradation pathway (6,7). Branched-chain α-keto acid decarboxylase (BCKDH-E1) is one of three enzymatic components in this complex (7). The α subunit of BCKDH-E1 (BCKDH-E1α) is critical for the regulation of BCKDH. Phosphorylation of BCKDH-E1α was shown to play a key role in regulating the enzymatic activity of this complex (7-9). Phosphorylation of BCKDH-E1α at Ser293 inactivates BCKDH (7,8). A significant elevation in plasma BCAA levels was reported to correlate with increased phosphorylation of BCKDH-E1α at Ser293 and suppressed BCKDH activity in the liver of diabetic mice (9).
- Bledsoe, R.K. et al. (1997) Biochim Biophys Acta 1339, 9-13.
- Suryawan, A. et al. (1998) Am J Clin Nutr 68, 72-81.
- Yoshikawa, R. et al. (2006) World J Gastroenterol 12, 5884-9.
- She, P. et al. (2007) Cell Metab 6, 181-94.
- Li, T. et al. (2017) Cell Metab 25, 374-385.
- Shin, A.C. et al. (2014) Cell Metab 20, 898-909.
- Lu, G. et al. (2009) J Clin Invest 119, 1678-87.
- Harris, R.A. et al. (1997) Adv Enzyme Regul 37, 271-93.
- Lian, K. et al. (2015) Diabetes 64, 49-59.
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