Tumor Necrosis Factor Alpha (TNFa) BioAssay™ ELISA Kit (Multi-species)

The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of TNFa in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids. Due to the 95% homology of the sequence among different species, the kit can be used to detect mouse and rat samples.

Sensitivity:
6.1pg/ml

Range:
15.6-1000pg/ml

Specificity:
This assay has high sensitivity and excellent specificity for detection of Tumor Necrosis Factor Alpha (TNFa). No significant cross-reactivity or interference between Tumor Necrosis Factor Alpha (TNFa) and analogues was observed.

Precision:
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Test Principle:
The microplate provided in this kit has been pre-coated with an antibody specific to TNFa. Standards or samples are then added to the appropriate microplate wells with a biotin-conjugated antibody specific to TNFa. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain TNFa, biotin-conjugated antibody and
enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of TNFa in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Kit Components:
96-well strip plate, 1x Plate
Standard, 2x vial
Detection Reagent A, 1×120ul
Detection Reagent B, 1×120ul
TMB Substrate, 1×9ml
Wash Buffer (30X), 1×20ml
Standard Diluent, 1×20ml
Assay Diluent A, 1×12ml
Assay Diluent B, 1×12ml
Stop Solution, 1×6ml

Storage and Stability:
Store microtiter strips, Standard, Detection Reagent A, Detection Reagent B and positive control at -20°C. After reconstitution of the postive control store at 4°C and use within 5 days. Store all the other components at 4°C. Unused kit is stable for 6 months after receipt. Once kit components are opened, it is highly recommended to use remaining reagents within 1 month provided this is within the expiration date of the kit. For maximum recovery of product, centrifuge the original vials after thawing and prior to removing the cap.