The Canine Adrenocorticotropic Hormone (ACTH) ELISA Kit is a highly sensitive and specific assay designed for the accurate detection of ACTH levels in canine serum, plasma, and cell culture supernatants. This kit offers reliable and reproducible results, making it suitable for a variety of research applications.ACTH is a hormone that plays a crucial role in the regulation of the adrenal glands and the production of cortisol.
Dysregulation of ACTH levels can lead to various endocrine disorders, making it an important biomarker for studying conditions such as Cushing's disease and Addison's disease in dogs.The Canine ACTH ELISA Kit provides researchers with a powerful tool for studying the role of ACTH in canine health and disease, paving the way for potential diagnostic and therapeutic advancements in veterinary medicine.
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 condition. 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 50µL standard or sample to each well.    And then add 50µL prepared Detection Reagent A immediately.    Shake and mix. Incubate 1 hour at 37°C
3.
Aspirate and wash 3 times
4.
Add 100µL prepared Detection Reagent B. Incubate 1 hour at 37°C
5.
Aspirate and wash 5 times
6.
Add 90µL Substrate Solution. Incubate 15-25 minutes at 37°C
7.
Add 50µL Stop Solution. Read at 450 nm immediately.
