Rat Tumor Necrosis Factor Alpha Elisa Kit For Tnf Alpha 48 Wells

Rat Tumor necrosis factor α,Cachectin,TNF,TNFSF2,TNF-α ELISA KIT

Materials provided with the kit

Principle

This ELISA kit uses Sandwich-ELISA as the method. The Microelisa stripplate provided in this kit has been pre-coated with an antibody specific to TNF-α . Standards or samples are added to the appropriate Microelisa stripplate wells and combined to the specific antibody. Then a Horseradish Peroxidase (HRP)- conjugated antibody specific for TNF-α is added to each Microelisa stripplate 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 thrombomodulin 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.

Notes:

• Store the kit at 4°C upon receipt.The kit should be equilibrated to room temperature before the assay. Remove any unneeded strips from TNF-α Antibody-Coated plate, reseal them in zip-lock foil and keep at 4°C.
• Precipitates may appear in concentrated washing buffer. Please heat the buffer to dissolve all the precipitates, which will not affect the results.
• Accurate pipette should be used to avoid experimental error. Samples should be added to the Microplate in less than 5 minutes. If a large number of samples are included, multiple channel pipette is recommended.
• Standard curve should be included in every assay. Replicate wells are recommended. If the OD value of the sample is greater than the first well of standards, please dilute the sample (n times) before test. When calculating the original thrombomodulin concentration, please multiply the total dilution factor (XnX5).
• In order to avoid cross-contamination, Microplate sealers are for one-time use only.
• Please keep Substrate away from light.
• All the operation should be accordance with the manufacturer's instructions strictly. The results determined by the Microtiter Plate Reader.
• All the samples, washing buffer and wastes should be treated as infectious agents.
• Reagents from different lots should not be mixed.

Known concentrations of TNF-a Standard and its corresponding reading OD is plotted on the log scale (x-axis) and the log scale (y-axis) respectively. The concentration of TNF-a in sample is determined by plotting the sample’s O.D. on the Y-axis. The original concentration is calculated by multiplying the dilution factor.