Immunoprecipitation of BCKDH-E1α from MCF7 extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is BCKDH-E1α (E4T3D) Rabbit mAb. Western blot analysis was performed using BCKDH-E1α (E4T3D) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as the secondary antibody.
Immunoprecipitation of Phospho-BCKDH-E1α (Ser293) from 293T extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-BCKDH-E1α (Ser293) (E2V6B) Rabbit mAb. Western blot analysis was performed using Phospho-BCKDH-E1α (Ser293) (E2V6B) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as the secondary antibody.
Western blot analysis of extracts from human liver, MCF7 and PANC-1 cell lines using BCKDH-E1α (E4T3D) Rabbit mAb.
Western blot analysis of extracts from various cell lines using Phospho-BCKDH-E1α (Ser293) (E2V6B) Rabbit mAb.
Western blot analysis of extracts from HeLa cells, untreated (-) or treated (+) with calf intestinal alkaline phosphatase (CIP)/λ phosphatase, using Phospho-BCKDH-E1α (Ser293) (E2V6B) Rabbit mAb (upper) or BCKDH-E1α (E4T3D) Rabbit mAb #90198 (lower).
PhosphoPlus® Duets from Cell Signaling Technology (CST) provide a means to assess protein activation status. Each Duet contains an activation-state and total protein antibody to your target of interest. These antibodies have been selected from CST's product offering based upon superior performance in specified applications.
Branched-chain amino acids (BCAAs) leucine, isoleucine, and valine are essential amino acids in mammals, but elevated levels of BCAAs have been implicated in cardiovascular and metabolic disorders (1). The branched-chain α-keto acid dehydrogenase complex (BCKDH) catalyzes the rate-limiting step in the BCAA degradation pathway (2,3). Branched-chain α-keto acid decarboxylase (BCKDH-E1) is one of three enzymatic components in this complex (3). 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 (3-5).
Phosphorylation of BCKDH-E1α at Ser293 inactivates BCKDH (3,4). 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 (5).
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