The Mouse ATPase H+/K+ Exchanging Alpha Polypeptide (ATP4A) ELISA Kit is specifically designed for the accurate detection of ATP4A levels in mouse serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring dependable and consistent results for a variety of research applications.ATP4A is an essential protein involved in ion transport and plays a crucial role in maintaining cellular pH balance. Dysfunction of ATP4A has been associated with various disease conditions, making it a valuable biomarker for studying diseases such as gastric ulcers, acid reflux, and other related disorders.
By accurately measuring ATP4A levels, researchers can gain valuable insights into the mechanisms underlying these diseases and potential therapeutic targets. The Mouse ATPase H+/K+ Exchanging Alpha Polypeptide (ATP4A) ELISA Kit provides a reliable tool for precise quantification of ATP4A levels, enabling researchers to advance their understanding of these important biological processes.
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)
83-100
91
EDTA plasma (n=5)
91-98
94
Heparin plasma (n=5)
90-99
94
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.
