The Canine Tumor Necrosis Factor Alpha (TNF-α) ELISA Kit is specifically designed for the accurate detection of TNF-α levels in canine serum, plasma, and cell culture supernatants. With high sensitivity and specificity, this kit ensures reliable and reproducible results, making it an essential tool for a variety of research applications related to inflammation, immune response, and cancer in canines.TNF-α is a key cytokine involved in the regulation of immune responses and inflammation. Abnormal levels of TNF-α have been linked to various diseases, including cancer, autoimmune disorders, and inflammatory conditions.
By accurately measuring TNF-α levels, researchers can gain valuable insights into the pathogenesis of these diseases and explore new therapeutic strategies.Overall, the Canine TNF-α ELISA Kit offers researchers a powerful tool for studying the role of TNF-α in canine health and disease, providing valuable information that can contribute to the development of new diagnostic and treatment options for canine patients.
Matrices listed below were spiked with certain level of recombinant the index and the recovery rates were calculated by comparing the measured value to the expected amount of the index in samples.
Matrix
Recovery range (%)
Average(%)
Serum (n=5)
80-102
91
EDTA plasma (n=5)
81-99
90
Heparin plasma (n=5)
80-89
84
Linearity:
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of the index and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample
1:2
1:4
1:8
1:16
Serum (n=5)
82-96%
83-98%
81-99%
93-101%
EDTA plasma (n=5)
88-101%
86-95%
90-102%
80-93%
Heparin plasma (n=5)
80-91%
82-90%
95-104%
79-95%
Intra-assay Precision:
Intra-Assay: CV <10%. 3 samples with low, middle and high level the index were tested 20 times on one plate, respectively.
Inter-assay Precision:
Inter-Assay: CV <12%. 3 samples with low, middle and high level the index were tested on 3 different plates, 8 replicates in each plate.
Stability:
The stability of ELISA kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage conditions. Note: To minimize unnecessary influences on the performance, operation procedures and lab conditions, especially room temperature, air humidity and incubator temperatures should be strictly regulated. It is also strongly suggested that the whole assay is performed by the same experimenter from the beginning to the end.
Step
Protocol
1.
Prepare all reagents, samples and standards
2.
Add 100µL standard or sample to each well. Incubate 2 hours at 37°C
3.
Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C
4.
Aspirate and wash 3 times
5.
Add 100µL prepared Detection Reagent B. Incubate 1 hour at 37°C
6.
Aspirate and wash 5 times
7.
Add 90µL Substrate Solution. Incubate 15-25 minutes at 37°C
8.
Add 50µL Stop Solution. Read at 450nm immediately.
