Sheep TNFa Kit: Ultra-High Tumor Necrosis Factor Alpha ELISA Detection
The Sheep TNFa Kit uses Sandwich enzyme immunoassay. The provided microtiter plate comes pre-coated with an antibody specific to Tumor Necrosis Factor Alpha (TNFa) .
Sheep TNFa Kit: Ultra-High Tumor Necrosis Factor Alpha ELISA Detection :
It is a specialized assay kit used for the detection and quantification of Tumor Necrosis Factor Alpha (TNF-α) in sheep samples. TNF-α is a pro-inflammatory cytokine that plays a critical role in immune responses, inflammation, and various disease processes.
The steps involved in using the Sheep TNFa Kit: Ultra-High Tumor Necrosis Factor Alpha ELISA Detection :
-Sample Preparation: Collect and prepare the sheep samples, such as serum, plasma, or cell culture supernatant, according to the kit's instructions. Ensure appropriate sample handling, storage, and dilution, if necessary.
-Plate Coating: Add the provided capture antibody specific to TNF-α to the wells of a microplate. Incubate the plate for a specific duration, usually 1-2 hours at room temperature or overnight at 4°C, to allow the capture antibody to bind to the TNF-α in the sample.
-Blocking: Block the remaining uncoated surfaces of the microplate wells to prevent non-specific binding. This is typically done by adding a blocking buffer, such as BSA or non-fat milk, and incubating the plate for a specified time.
-Sample and Standard Addition: Add the prepared sheep samples and a set of known TNF-α standards with different concentrations to the respective wells of the microplate. This allows the standards to generate a calibration curve for quantification.
-Antibody Incubation: Add a detection antibody specific to TNF-α, labeled with an enzyme or a fluorophore, to each well. Incubate the plate for a specific duration to allow the detection antibody to bind to the captured TNF-α.
-Washing: Wash the microplate wells multiple times with a suitable wash buffer to remove any unbound substances and reduce background noise.
-Substrate Addition: Add a substrate solution that reacts with the enzyme or fluorophore conjugated to the detection antibody. This reaction generates a detectable signal that is directly proportional to the amount of TNF-α present in the samples.
-Color Development: Incubate the plate for a specified time to allow the color to develop. The intensity of the color is proportional to the concentration of TNF-α in the samples.
-Stop Reaction: Add a stop solution to halt the color development and stabilize the reaction.
-Measurement: Using a microplate reader, measure the absorbance or fluorescence of each well at the appropriate wavelength. This provides quantitative data for the TNF-α concentration in the samples.
-Data Analysis: Analyze and interpret the results based on the standard curve and the sample values, following the kit's instructions. Calculate the concentration of TNF-α in the samples using the standard curve.