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Figure 2. The relationship between the protein concentration of lysates from control NIH/3T3 cells or LY294002-treated NIH/3T3 cells and the absorbance at 450 nm is shown. NIH/3T3 cells (60-70% confluent) were treated with LY294002 #9901 (16 μM, 48 hr) and then lysed.Learn more about how we get our images
Figure 1. Treatment of NIH/3T3 cells with LY294002 decreases expression of FASN as detected by the PathScan® Total Fatty Acid Synthase Sandwich ELISA Kit #7689. NIH/3T3 cells (60-70% confluent) were treated with LY294002 #9901 (16 μM, 48 hr). The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blot using Fatty Acid Synthase (C20G5) Rabbit mAb #3180 is shown in the bottom figure.Learn more about how we get our images
|Product Includes||Volume||Solution Color|
|FASN Rabbit Ab Coated Microwells||96 tests|
|Fatty Acid Synthase (C20G5) Rabbit Detection mAb (Biotinylated)||1 ea||Green (Lyophilized)|
|HRP-Linked Streptavidin (ELISA Formulated)||1 ea||Red (Lyophilized)|
|Detection Antibody Diluent 3||11 ml||Green|
|HRP Diluent||11 ml||Red|
|TMB Substrate 7004||11 ml|
|STOP Solution 7002||11 ml|
|Sealing Tape||2 ea|
|Cell Lysis Buffer (10X) 9803||15 ml|
|ELISA Sample Diluent||25 ml||Blue|
|ELISA Wash Buffer (20X) 9801||25 ml|
NOTE: Prepare solutions with purified water.
*NOTE: Some PathScan® ELISA Kits may include HRP-Linked Streptavidin in place of HRP-Linked Antibody.
NOTE: Initial color of positive reaction is blue, which changes to yellow upon addition of STOP Solution.
posted November 2013
The PathScan® Total Fatty Acid Synthase Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of fatty acid synthase protein (FASN). A FASN Rabbit Antibody has been coated onto the microwells. After incubation with cell lysates, FASN is captured by the coated antibody. Following extensive washing, a biotinylated FASN Rabbit Detection Antibody is added to detect the captured FASN protein. HRP-linked streptavidin is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for the developed color is proportional to the quantity of FASN.
Antibodies in kit are custom formulations specific to kit.
This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Human, Mouse, Monkey
Fatty acid synthase (FASN) catalyzes the synthesis of long-chain fatty acids from acetyl-CoA and malonyl-CoA. FASN is active as a homodimer with seven different catalytic activities and produces lipids in the liver for export to metabolically active tissues or storage in adipose tissue. In most other human tissues, FASN is minimally expressed since they rely on circulating fatty acids for new structural lipid synthesis (1).
According to the research literature, increased expression of FASN has emerged as a phenotype common to most human carcinomas. For example in breast cancer, immunohistochemical staining showed that the levels of FASN are directly related to the size of breast tumors (2). Research studies also showed that FASN is highly expressed in lung and prostate cancers and that FASN expression is an indicator of poor prognosis in breast and prostate cancer (3-5). Furthermore, inhibition of FASN is selectively cytotoxic to human cancer cells (5). Thus, increased interest has focused on FASN as a potential target for the diagnosis and treatment of cancer as well as metabolic syndrome (6,7).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. PathScan is a trademark of Cell Signaling Technology, Inc.
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