TNF alpha ELISpot Assay Kit (Rat)

ELISA Kit

Wet Ice

TNF

235

acts as a cytokine; binds TNF receptors; plays a role in regulation of cell proliferation, induction of apoptosis, and inflammatory response [RGD, Feb 2006]

NP_036807.1

Assay Methodology: Quantitative Sandwich ELISpot Immunoassay

Aviva's Rat TNF alpha ELISpot Kit is based on standard sandwich ELISA methodology. A capture antibody highly specific for the analyte of interest is coated to the wells of a PVDF bottomed 96 well microtitre plate either during kit manufacture or in the laboratory. The plate is then blocked to minimise any non-antibody dependent unspecific binding and washed. Cell suspension and stimulant are added and the plate incubated allowing the specific antibodies to bind any analytes produced. Cells are then removed by washing prior to the addition of Biotinylated detection antibodies which bind to the previously captured analyte. Enzyme conjugated streptavidin is then added binding to the detection antibodies. Following incubation and washing substrate is then applied to the wells resulting in coloured spots which can be quantified using appropriate analysis software or manually using a microscope.