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Analysis of Fas Ligand/TNFSF6 by ELISA (CAT#: STEM-MB-1532-LGZ)

Introduction

Fas ligand, also known as TNFSF6, FasL, or CD95L, is a member of the tumor necrosis factor (TNF) family. It is a homologous trimer type II transmembrane protein expressed on cytotoxic T lymphocytes. Extracellular matrix metalloproteinase MMP-7 cleaved membrane-bound FasL at conserved cleavage sites to produce soluble Fas ligands. Fas ligand/receptor interactions play an important role in regulating the immune system and cancer progression. It generates signals through trimerization of FASRs, which usually leads to apoptosis or death. Apoptosis induced by Fas-Fas ligand binding plays a critical role in the regulation of the immune system. Its functions include T cell homeostasis, cytotoxic T cell activity, immune privilege, maternal tolerance, and tumor counterattack. Defective Fas mediated apoptosis can lead to tumor development and drug resistance in existing tumors. Mutations in Fas genes are associated with an autoimmune lymphoproliferative syndrome (ALPS).




Principle

Enzyme-linked immunosorbent assay (ELISA) is an enzyme-labeled solid phase immunoassay technique. Its basic principle is to bind the antigen (or antibody) to the solid phase carrier, and the antigen (or antibody) and a certain enzyme link to enzyme labeled antigen (or antibody). During detection, the sample to be tested and the enzymic antigen (or antibody) react with the antigen (or antibody) on the solid phase carrier according to certain procedures, and then remove the unreacted part by washing method. After adding the substrate, the substrate is catalyzed by the enzyme on the solid phase carrier to produce colored substances. Through qualitative or quantitative detection of the amount of colored products, the content of the substance to be measured in the sample can be determined.

Applications

Oncology & Cancer, Immunology/Inflammation

Procedure

1. Add standards or samples to each well and incubate.
2. Pour off the liquid in the well, biotinylated antibody working solution and incubate.
3. Add enzyme conjugate working solution and incubate.
4. Add substrate TMB and incubate.
5. Add stop solution and measure OD value.
6. Calculation of results.

Materials

• Sample Type: Serum, plasma, cell culture supernatant and other biological samples

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