Purpose

Our Mouse Tumor necrosis factor Alpha, TNF-α ELISA Kit is to assay TNF-α levels in Mouse serum, plasma, culture media or any biological fluid.

Principle

This ELISA kit uses Sandwich-ELISA as the method. The Micro Elisa strip plate provided in this kit has been pre-coated with an antibody specific to TNF-α. Standards or samples are added to the appropriate Micro Elisa strip plate wells and combined to the specific antibody. Then a Horseradish Peroxidase (HRP)-conjugated antibody specific for TNF-α is added to each Micro Elisa strip plate well and incubated. Free components are washed away. The TMB substrate solution is added to each well. Only those wells that contain TNF-α and HRP conjugated TNF-α antibody will appear blue in color and then turn yellow after the addition of the stop solution. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm. The OD value is proportional to the concentration of TNF-α. You can calculate the concentration of TNF-α in the samples by comparing the OD of the samples to the standard curve.

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Mouse TNF-α were tested 20 times on one plate, respectively

Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Mouse TNF-α were tested on 3 different plates, 8 replicates in each plate.

CV(%) = SD/meanX100

Intra-Assay: CV<10%

Inter-Assay: CV<12%