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34995
Branched-Chain Amino Acid Metabolism Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Branched-Chain Amino Acid Metabolism Antibody Sampler Kit #34995

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Branched-Chain Amino Acid Metabolism Antibody Sampler Kit: Image 1
Western blot analysis of extracts from various cell lines using Phospho-BCKDH-E1α (Ser293) (E2V6B) Rabbit mAb.
Branched-Chain Amino Acid Metabolism Antibody Sampler Kit: Image 2
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Branched-Chain Amino Acid Metabolism Antibody Sampler Kit: Image 3
Western blot analysis of extracts from MCF7, COS-7, and Vero cell lines using BCAT2 (D8K3O) Rabbit mAb.
Branched-Chain Amino Acid Metabolism Antibody Sampler Kit: Image 4
Western blot analysis of Jurkat, MOLT-4, and Neuro-2a cells using BCAT1 (D6D4K) Rabbit mAb.
Branched-Chain Amino Acid Metabolism Antibody Sampler Kit: Image 5
Western blot analysis of extracts from human liver, MCF7 and PANC-1 cell lines using BCKDH-E1α (E4T3D) Rabbit mAb.
Branched-Chain Amino Acid Metabolism Antibody Sampler Kit: Image 6
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).
Branched-Chain Amino Acid Metabolism Antibody Sampler Kit: Image 7

Immunohistochemical analysis of paraffin-embedded human ductal carcinoma of the breast using BCAT2 (D8K3O) Rabbit mAb.
Branched-Chain Amino Acid Metabolism Antibody Sampler Kit: Image 8
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.
Branched-Chain Amino Acid Metabolism Antibody Sampler Kit: Image 9
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.
Branched-Chain Amino Acid Metabolism Antibody Sampler Kit: Image 10
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using BCAT2 (D8K3O) Rabbit mAb.
Branched-Chain Amino Acid Metabolism Antibody Sampler Kit: Image 11
Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using BCAT2 (D8K3O) Rabbit mAb.
Branched-Chain Amino Acid Metabolism Antibody Sampler Kit: Image 12

Immunohistochemical analysis of paraffin-embedded human serous papillary carcinoma of the ovary using BCAT2 (D8K3O) Rabbit mAb.
Branched-Chain Amino Acid Metabolism Antibody Sampler Kit: Image 13
Immunohistochemical analysis of paraffin-embedded SCLC-21H cell pellet (left, high-expressing) or HepG2 cell pellet (right, low-expressing) using BCAT2 (D8K3O) Rabbit mAb.
Branched-Chain Amino Acid Metabolism Antibody Sampler Kit: Image 14
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using BCAT2 (D8K3O) Rabbit mAb (left) compared to concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
To Purchase # 34995T
Product # Size Price
34995T
1 Kit  (6 x 20 microliters) $ 336

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
BCAT1 (D6D4K) Rabbit mAb 88785 20 µl
  • WB
H M 43 Rabbit IgG
BCAT2 (D8K3O) Rabbit mAb 79764 20 µl
  • WB
  • IP
  • IHC
H Mk 39 Rabbit IgG
BCKDH-E1α (E4T3D) Rabbit mAb 90198 20 µl
  • WB
  • IP
H M R 49 Rabbit IgG
Phospho-BCKDH-E1α (Ser293) (E2V6B) Rabbit mAb 40368 20 µl
  • WB
  • IP
H M R Mk 49 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

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).
  1. Bledsoe, R.K. et al. (1997) Biochim Biophys Acta 1339, 9-13.
  2. Suryawan, A. et al. (1998) Am J Clin Nutr 68, 72-81.
  3. Yoshikawa, R. et al. (2006) World J Gastroenterol 12, 5884-9.
  4. She, P. et al. (2007) Cell Metab 6, 181-94.
  5. Li, T. et al. (2017) Cell Metab 25, 374-385.
  6. Shin, A.C. et al. (2014) Cell Metab 20, 898-909.
  7. Lu, G. et al. (2009) J Clin Invest 119, 1678-87.
  8. Harris, R.A. et al. (1997) Adv Enzyme Regul 37, 271-93.
  9. Lian, K. et al. (2015) Diabetes 64, 49-59.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
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