Human TNF alpha ELISpot Kit (A334659)

Human TNF alpha ELISpot Kit

Human TNF alpha ELISpot Kit is a ready-to-use ELISpot assay for the in vitro detection and quantification of TNF alpha-secreting human PBMCs at the single-cell level.

sandwich ELISA

The Human TNF alpha ELISpot Kit (A334659) employs the double-antibody sandwich ELISpot technique for the detection of TNF alpha-secreting cells. A capture antibody specific for TNF alpha has been pre-coated onto a 96-well microplate. Test cells, such as PBMCs, are added into the wells, and upon stimulation with a cell stimulant, TNF alpha secreted by individual cells is captured by the immobilized antibody. A biotinylated anti-TNF alpha detection antibody, which also binds the captured cytokine, and Streptavidin-HRP, which binds the biotinylated detection antibody, are then added, and the plate is incubated. Following incubation, unbound detection antibody and Streptavidin-HRP are removed by washing, and AEC substrate solution is added. Colored spots develop at the sites of cytokine secretion in proportion to the number of TNF alpha-producing cells. The intensity and number of spots can then be quantified using an ELISpot reader. This ready-to-use kit includes pre-coated plates, detection antibody, Streptavidin-HRP, AEC coloring system, dilution buffer, cell stimulant, and washing buffer. For research use only. Not for diagnostic purposes.

Biotinylated Antibody 100µl 1 vial; Streptavidin-HRP 100µl 1 vial; Cell Stimulant 50 T 1 bottle; Dilution Buffer R (1×) 10mL 2 bottles; Washing Buffer (50×) 15mL 1 bottle; AEC Dilution 10mL 1 bottle; AEC Solution I (20×) 500µl 1 vial; AEC Solution II (20×) 500µl 1 vial; AEC Solution III (200×) 50µl 1 vial; Pre-coated PVDF Plate/1 plate.

Precoated PVDF Microplate

ELISpot Plate Reader or Image Capture and Quantification. Spots on the membrane are counted, with each spot corresponding to one secreting cell, providing a quantitative measure of cellular activity.

Peripheral blood mononuclear cells (PBMCs) in single-cell suspension, capable of cytokine secretion.

Assay time: 3 hours. Sample incubation time: 16-24 hours.

This ELISpot Kit specifically targets Human TNF alpha.

Human

Intra-assay CV: < 15%, Inter-assay CV: < 20%

Shipped at 4°C. The kit should be stored at 2-8°C. Shelf life is 12 months from the date of receipt.

Cachectin, TNF, TNF-a, TNF-alpha, TNFA, TNFSF2, Tumor necrosis factor, Tumor necrosis factor ligand superfamily member 2

Information online is representative. Always refer to the Product Manual inside the kit.