The Rat Pyruvate Kinase (PK) Liver and RBC ELISA Kit is a reliable and sensitive tool for measuring pyruvate kinase levels in rat liver and red blood cells. This kit offers accurate detection in serum, plasma, and cell culture supernatants, allowing for precise and reproducible results in a variety of research applications.Pyruvate kinase is an essential enzyme involved in glycolysis, playing a critical role in energy metabolism and cellular function. Abnormalities in pyruvate kinase levels have been associated with a range of diseases, making this kit valuable for studying metabolic disorders, liver diseases, and other conditions where pyruvate kinase activity is important.
With its high sensitivity and specificity, the Rat Pyruvate Kinase Liver and RBC ELISA Kit provides researchers with a valuable tool for exploring the role of pyruvate kinase in health and disease, offering insight into potential therapeutic targets and diagnostic markers.
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.
