Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Total Acetyl-CoA Carboxylase Sandwich ELISA Kit #7996


No. Size Price
7996C 1 Kit ( 96 assays ) ¥6,345.00 现货查询 购买询价
Product Includes Volume Solution Color
ELISA Sample Diluent 25 ml Blue
Anti-rabbit IgG, HRP-linked Antibody (ELISA Formulated) 1 ea Red (Lyophilized)
Detection Antibody Diluent 11 ml Green
HRP Diluent 11 ml Red
Acetyl-CoA Carboxylase 1 Rabbit Detection Ab 1 ea Green (Lyophilized)
STOP Solution #7002 11 ml Colorless
TMB Substrate #7004 11 ml Colorless
PathScan® Sandwich ELISA Lysis Buffer (1X) #7018 30 ml Colorless
ELISA Wash Buffer (20X) 25 ml Colorless
ACC Mouse Ab Coated Microwells 96 tests
Sealing Tape 2 sheets

Specificity / Sensitivity

PathScan® Total Acetyl-CoA Carboxylase Sandwich ELISA Kit #7996 detects endogenous levels of total ACC protein (see Figure 1). The kit sensitivity is shown in Figure 2.

PathScan® Total Acetyl-CoA Carboxylase Sandwich ELISA Kit #7996用于检测内源性乙酰辅酶A合成酶(ACC)总蛋白水平(见图1)。试剂盒敏感性见图2。


The PathScan® Total Acetyl-CoA Carboxylase (ACC) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of ACC. An ACC mouse antibody has been coated onto the microwells. After incubation with cell lysates, ACC (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, an ACC rabbit detection antibody is added to detect captured ACC protein. Anti-rabbit IgG, HRP-linked Antibody #7074 is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of total ACC.

PathScan® Total Acetyl-CoA Carboxylase (ACC) Sandwich ELISA Kit是一种固相三明治酶联免疫试验(ELISA)试剂盒,用于检测内源性ACC。首先,小鼠ACC抗体涂在微孔中表面,然后与细胞裂解液孵育,裂解液中的ACC蛋白(磷酸化的和非磷酸化的)被捕获到涂层抗体上。将微孔洗脱后,加入兔ACC检测抗体,用于检测被捕获的ACC蛋白。然后加入抗兔IgG抗体,即辣根过氧化物酶耦联抗体( HRP-linked Antibody #7074),用于检测已经结合的检测抗体。最后加入辣根过氧化物酶的底物,四甲基联苯胺(TMP),加入底物后发生显色反应。ACC的数量与显色的强度成正比,因此可以借此对ACC定量。

ELISA - Western correlation

ELISA - Western correlation

Figure 1. Treatment of Hep G2 cells with H2O2 stimulates phosphorylation of ACC at Ser79, detected by the PathScan® Phospho-ACC (Ser79) Sandwich ELISA Kit #7986, but does not affect the levels of total ACC detected by PathScan® Total ACC Sandwich ELISA Kit #7996. Hep G2 cells (80-90% confluent) were treated 10 mM hydrogen peroxide for 10 minutes and lysed with #7018. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blots using Acetyl-CoA Carboxylase (C83B10) Rabbit mAb #3676 (left panel) or Phospho-Acetyl-CoA Carboxylase (Ser79) Antibody #3661 (right panel) are shown in the bottom figure.

图1,用过氧化氢处理Hep G2细胞会增加ACC在Ser79位点的磷酸化水平,检测试剂盒为PathScan® Phospho-ACC (Ser79) Sandwich ELISA Kit #7986。但不影响ACC总蛋白水平,检测试剂盒为PathScan® Total ACC Sandwich ELISA Kit #7996。Hep G2用10 mM过氧化氢处理10分钟,然后用#7018裂解。上图所示为450nm处的吸光度值,下图所示为相应的Western blot检测,所用抗体为Acetyl-CoA Carboxylase (C83B10) Rabbit mAb兔单抗 #3676(左),Phospho-Acetyl-CoA Carboxylase (Ser79) Antibody #3661(右)。



Figure 2. The relationship between the protein concentration of lysates from untreated and H2O2-treated Hep G2 cells and the absorbance at 450 nm using the PathScan® Total ACC Sandwich ELISA Kit #7996 is shown.

图2,Hep G2细胞裂解液中蛋白浓度与450钠米处吸光度的关系图,所用细胞未处理或用过氧化氢处理,所用试剂盒为PathScan® Total ACC Sandwich ELISA Kit #7996


Acetyl-CoA carboxylase (ACC) catalyzes the pivotal step of the fatty acid synthesis pathway. The 265 kDa ACCα (ACC1) is the predominant isoform found in liver, adipocytes, and mammary gland, while the 280 kDa ACCβ (ACC2) is the major isoform in skeletal muscle and heart (1). Phosphorylation by AMPK at Ser79 or by PKA at Ser1200 inhibits the enzymatic activity of ACC (2). ACC is a potential target of anti-obesity drugs (3,4).

乙酰辅酶A合成酶(ACC)用于催化脂肪酸合成途径中的关键步骤。分子量为265 kDa的ACCα,是肝脏,脂肪细胞和乳腺中ACC的主要形式,而在骨胳肌和心脏中,其主要形式为280 kDa的ACCβ(1)。ACC可以在Ser79被AMPK磷酸化,或者在Ser1200被PKA磷酸化,这些位点的磷酸化可以抑制ACC的酶活性(2)。ACC也是减肥药物的一个潜在靶点(3,4)。

  1. Castle, J.C. et al. (2009) PLoS One 4, e4369.
  2. Kreuz, S. et al. (2009) Diabetes Metab Res Rev 25, 577-86.
  3. Ha, J. et al. (1994) J Biol Chem 269, 22162-8.
  4. Abu-Elheiga, L. et al. (2001) Science 291, 2613-6.
  5. Levert, K.L. et al. (2002) J Biol Chem 277, 16347-50.

Application References

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Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

PathScan is a trademark of Cell Signaling Technology, Inc.

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