Revision 1

#34995Store at -20C

1 Kit

(6 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
BCAT1 (D6D4K) Rabbit mAb 88785 20 µl 43 kDa Rabbit IgG
BCAT2 (D8K3O) Rabbit mAb 79764 20 µl 39 kDa Rabbit IgG
BCKDH-E1α (E4T3D) Rabbit mAb 90198 20 µl 49 kDa Rabbit IgG
Phospho-BCKDH-E1α (Ser293) (E2V6B) Rabbit mAb 40368 20 µl 49 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

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.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibodies.

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.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    使用に関する制限

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    Revision 1
    #34995

    Branched-Chain Amino Acid Metabolism Antibody Sampler Kit

    Branched-Chain Amino Acid Metabolism Antibody Sampler Kit: Image 1 Expand Image
    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 Expand Image
    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 Expand Image
    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 Expand Image
    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 Expand Image
    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 Expand Image
    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 Expand Image
    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 Expand Image
    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 Expand Image
    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 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using BCAT2 (D8K3O) Rabbit mAb.
    Branched-Chain Amino Acid Metabolism Antibody Sampler Kit: Image 11 Expand Image
    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 Expand Image

    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 Expand Image
    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 Expand Image
    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).