Mouse TNF alpha ELISA Kit

Product details

Mouse TNF alpha ELISA Kit ab100747 is a sandwich ELISA to measure Mouse TNF alpha in plasma, cell culture supernatant, serum free media with a sensitivity of 60 pg/ml.

How the assay works

This assay employs an antibody specific for Mouse TNF alpha coated on a 96-well plate. Standards and samples are pipetted into the wells and TNF alpha present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Mouse TNF alpha antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of TNF alpha bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
Mouse TNFa levels are extremely low, and as such we expect "normal" levels to be undetectable when using our kit on plasma samples.
We have not been able to detect endogenous Mouse TNF alpha in normal serum with ab100747, only in serum spiked with Mouse TNF alpha.

Assay Specificity

Our ELISA kits are rigorously validated to ensure the highest level of consistency and reproducibility. Please check the protocol booklet for more details

Mouse TNF alpha ELISA Kit ab100747 protocol summary

1. Add standard or sample to each well used. Incubate at room temperature
2. Wash and add prepared biotinylated antibody to each well. Incubate at room temperature.
3. Wash and add prepared streptavidin solution. Incubate at room temperature.
4. Add TMB Development Solution to each well. Incubate at room temperature.
5. Add Stop Solution to each well. Read at 450nm immediately.