Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Total Fatty Acid Synthase Sandwich ELISA Kit #7689

When ordering five or more kits, please contact us for processing time and pricing at sales@cellsignal.com.

Kit Includes Volume Solution Color
FASN Rabbit Ab Coated Microwells 96 tests
Biotinylated FASN Rabbit Detection Ab 11 ml Green
Streptavidin-HRP 11 ml Red
TMB Substrate #7004 11 ml Colorless
STOP Solution #7002 11 ml Colorless
Sealing Tape 2 sheets
Cell Lysis Buffer (10X) #9803 15 ml Yellowish
ELISA Sample Diluent 25 ml Blue
ELISA Wash Buffer (20X) 25 ml Colorless

Note: 12 8-well modules –Each module is designed to break apart for 8 tests.
Note: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).

Species Cross-Reactivity

H M Mk

Reactivity Key:  H=Human  M=Mouse  Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Description

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.

Specificity / Sensitivity

This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

ELISA

ELISA

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.

ELISA - Western correlation

ELISA - Western correlation

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.

Background

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).

  1. Katsurada, A. et al. (1990) Eur J Biochem 190, 427-33.
  2. Wells, W.A. et al. (2006) Breast Cancer Res Treat 98, 231-40.
  3. Kawamura, T. et al. (2005) Pathobiology 72, 233-240.
  4. Shah, U.S. et al. (2006) Hum Pathol 37, 401-409.
  5. Kuhajda, F.P. (2000) Nutrition 16, 202-8.
  6. Tian, W.X. (2006) Curr Med Chem 13, 967-977.
  7. Kusunoki, J. et al. (2006) Endocrine 29, 91-100.

Application References

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Protocols

Companion Products

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

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