Product Pathways - PathScan ELISA
PathScan® Total Acetyl-CoA Carboxylase Sandwich ELISA Kit #7996
|7996S||1 Kit (96 assays)||---||In Stock||---|
|7996||carrier free and custom formulation / quantity||email request|
When ordering five or more kits, please contact us for processing time and pricing at email@example.com.
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|Kit Includes||Volume||Solution Color|
|ACC Mouse Ab Coated Microwells||96 assays|
|ACC Rabbit Detection Ab||11 ml||Green|
|Anti-rabbit IgG, HRP-linked Antibody||11 ml||Red|
|TMB Substrate #7004||11 ml||Colorless|
|STOP Solution #7002||11 ml||Colorless|
|Sealing Tape||2 sheets|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|ELISA Sample Diluent||25 ml||Blue|
|PathScan® Sandwich ELISA Lysis Buffer (1X) #7018||30 ml||Colorless|
Note: 12 8-well modules – Each module is designed to break apart for 8 tests.
Storage: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).
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.
Antibodies in kit are custom formulations specific to kit.
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.
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.
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.
Acetyl-CoA carboxylase (ACC) catalyzes the carboxylation of acetyl-CoA to malonyl-CoA (1). It is the key enzyme in the biosynthesis and oxidation of fatty acids (1). In rodents, the 265 kDa ACC1 (ACCα) form is primarily expressed in lipogenic tissues, while 280 kDa ACC2 (ACCβ) is the main isoform in oxidative tissues (1,2). However, in humans, ACC2 is the predominant isoform in both lipogenic and oxidative tissues (1,2). Phosphorylation by AMPK at Ser79 or by PKA at Ser1200 inhibits the enzymatic activity of ACC (3). ACC is a potential target of anti-obesity drugs (4,5).
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PathScan® is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.