The Mouse Apoptotic Peptidase Activating Factor 1 (APAF1) ELISA Kit is specifically designed for the precise measurement of APAF1 levels in mouse samples, including serum, plasma, and cell culture supernatants. This kit offers exceptional sensitivity and specificity, ensuring accurate and reproducible results for various research applications.APAF1 is a key regulator of apoptosis, playing a critical role in the activation of caspases and the initiation of the apoptotic pathway. Dysregulation of APAF1 expression has been linked to various diseases, including cancer, neurodegenerative disorders, and cardiovascular diseases, making it a valuable biomarker for understanding these conditions and potentially developing therapeutic interventions.
The Mouse APAF1 ELISA Kit is an essential tool for researchers studying apoptosis, cell death, and related pathways in mouse models. Its reliable performance and ease of use make it a valuable asset for advancing research in these areas and uncovering new insights into the mechanisms of disease.
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)
81-100
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.
