Mouse TNF-alpha ELISPOT Kit
组分（Materials Provided）
产品概述（Product Overview）
Tumor necrosis factor-alpha (TNF-α) is a multifunctional Th1 cytokine and one of the most important inflammatory cytokines. It is produced by macrophages during inflammation and can also be activated by the endotoxin lipopolysaccharide (LPS). The Mouse TNF-α ELISpot assay is designed for the detection of mouse TNF-α secreting cells at the single cell level and can be used to quantitate the frequency of mouse TNF-α secreting cells. ELISpot assays are highly reproducible and sensitive, and can be used to measure responses with frequencies well below 1 in 100,000. ELISpot assays do not require prior in vitro expansion of T cells and they are suitable for high-throughput analysis using only small volumes of primary cells. As such, ELISpot assays are useful tools for research in vaccine development, cytokine secretion and the monitoring of various clinical trials.
应用说明（Application）
This kit is used to detect TNF-alpha at the cellular level.
It is for research use only.
重构方法（Reconstitution）
Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.
存储（Storage）
1. Unopened kit should be stored at 2℃-8℃ upon receiving.
2. Find the expiration date on the outside packaging and do not use reagents past their expiration date.
3. The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.
原理（Assay Principles）
ELISpot assays employ the quantitative sandwich enzyme-linked immunosorbent assay (ELISA) technique. The Kit consists of Pre-coated Anti-TNF-α Antibody Microplate, Positive Stimulus, Biotin-Anti-TNF-α Antibody, Streptavidin-HRP, AEC and buffers.
Your experiment will include 6 simple steps:
a) A monoclonal antibody specific for mouse TNF-α has been pre-coated onto a polyvinylidene difluoride (PVDF)-backed microplate.
b) Appropriately stimulated cells are pipetted into the wells and the microplate is placed into a humidified 37 °C CO2 incubator for a specified period of time.During this incubation period, the immobilized antibody in the immediate vicinity of the secreting cells binds secreted TNF-α.
c) After washing away any cells and unbound substances, a biotinylated antibody specific for mouse TNF-α is added to the wells. Following a wash to remove any unbound biotinylated antibody, HRP conjugated to streptavidin is added.
d) Wash the plate and add the Streptavidin-HRP diluted by Dilution Buffer to the plate.
e) Wash the plate and add AEC. A reddish-brown colored precipitate forms and appears as spots at the sites of cytokine localization, with each individual spot representing an individual TNF-α secreting cell.
f) The spots can be counted with an automated ELISpot reader system or manually using a stereomicroscope.
关键字： TNF-alpha;TNF-alpha试剂盒;TNF-alpha ELISPOT试剂盒;ACRO;百普赛斯;
百普赛斯集团ACROBiosystems Group（股票代码：301080）是成立于2010年的跨国生物科技公司，是为全球生物医药、健康产业领域提供关键生物试剂产品及解决方案的行业平台型基石企业。2021年在创业板上市。百普赛斯集团业务遍布全球，横跨亚洲、北美洲、欧洲，在中国、美国、瑞士等12个城市设有办公室、研发中心及生产基地。目前累计服务客户超6000家，与全球Top 20医药企业均建立了长期、稳定的合作伙伴关系。集团旗下拥有品牌ACROBiosystems百普赛斯、bioSeedin柏思荟、Condense Capital垦拓资本和ACRODiagnostics百斯医学等。