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EK000071-EK0336: Mouse FAS ELISA Kit
Range 31.2pg/ml-2000pg/ml
Sensitivity < 3 pg/ml
Specificity: no detectable cross-reactivity with any other cytokine.
Application: for quantitative detection of mouse FAS in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
Fas, also known as APO-1, CD95 and TNFRSF6, is a member of the nerve growth factor (NGF)/tumour necrosis factor (TNF) receptor superfamily and mediates apoptosis.1 The nucleotide sequence of the cDNAs reveales that the molecule coding for the Fas antigen determinant is a 319 amino acid polypeptide with a single transmembrane domain. The extracellular domain is rich in cysteine residue, and shows a similarity to that of human tumor necrosis factor receptors, human nerve growth factor receptor, and human B cell antigen CD40.2 The APO-1 antigen as defined by the mouse monoclonal antibody anti-APO-1 is previously found to be expressed on the cell surface of activated human T and B lymphocytes and a variety of malignant human lymphoid cell lines. The APO-1 antigen is found to be a membrane glycoprotein of 48-kDa.3 Fas antigen is expressed and functional on papillary thyroid cancer cells and this may have potential therapeutic significance.4 Fas can play a role as an inducer of both neurite growth in vitro and accelerates recovery after nerve injury in vivo.5 The FAS and FASL triggered apoptosis pathway plays an important role in human carcinogenesis.6 The standard product used in this kit is FAS extracellular sections (1-173aa), in addition to the soluble human IgG1 Fc domain. The molecular mass is 45KDa.
Principle
The ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. The target specific antibodies are precoated onto 96-well plates. The target from the sample is bound to the microwell. The biotinylated target specific detection antibodies are added to the microwells and followed by washing with the PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex is added and unbound conjugates are washed away with the PBS or TBS buffer. TMB, the HRP (horseradish peroxidase) substrate, is used to visualize color change resulting from the HRP enzymatic reaction. TMB is catalyzed by HRP to produce the blue color. The color changes into yellow after the acidic stop solution is added. The density of the yellow color is proportional to the target amount from the sample captured in the microwells.
Reference
1. Dhein, J.; Walczak, H.; Baumler, C.; Debatin, K.-M.; Krammer, P. H. Autocrine T-cell suicide mediated by APO-1/(Fas/CD95). Nature 373: 438-441, 1995.
2. Itoh, N.; Yonehara, S.; Ishii, A.; Yonehara, M.; Mizushima, S.-I.; Sameshima, M.; Hase, A.; Seto, Y.; Nagata, S. The polypeptide encoded by the cDNA for human cell surface antigen Fas can mediate apoptosis. Cell 66: 233-243, 1991.
3. Oehm, A.; Behrmann, I.; Falk, W.; Pawlita, M.; Maier, G.; Klas, C.; Li-Weber, M.; Richards, S.; Dhein, J.; Trauth, B. C.; Ponstingl, H.; Krammer, P. H. Purification and molecular cloning of the APO-1 cell surface antigen, a member of the tumor necrosis factor/nerve growth factor receptor superfamily: sequence identity with the FAS antigen. J. Biol. Chem. 267: 10709-10715, 1992.
4. Arscott, P. L.; Stokes, T.; Myc, A.; Giordano, T. J.; Thompson, N. W.; Baker, J. R., Jr. Fas (CD95) expression is up-regulated on papillary thyroid carcinoma. J. Clin. Endocr. Metab. 84: 4246-4252, 1999.
5. Desbarats, J.; Birge, R. B.; Mimouni-Rongy, M.; Weinstein, D. E.; Palerme, J.-S.; Newell, M. K. Fas engagement induces neurite growth through ERK activation and p35 upregulation. Nature Cell Biol. 5: 118-125, 2003.
6. Zhang, X.; Miao, X.; Sun, T.; Tan, W.; Qu, S.; Xiong, P.; Zhou, Y.; Lin, D. Functional polymorphisms in cell death pathway genes FAS and FASL contribute to the risk of lung cancer. J. Med. Genet. 42: 479-484, 2005.
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