爱帕琳肽12(AP12)检测试剂盒说明书

ELISA Kit for Apelin 12 (AP12)

FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!

Specificity

This assay has high sensitivity and excellent specificity for detection of Apelin 12 (AP12).
No significant cross-reactivity or interference between Apelin 12 (AP12) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Apelin 12 (AP12) and the recovery rates were calculated by comparing the measured value to the expected amount of Apelin 12 (AP12) in samples.

Matrix	Recovery range (%)	Average(%)
serum(n=5)	98-105	101
EDTA plasma(n=5)	97-105	101
heparin plasma(n=5)	87-96	90

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Apelin 12 (AP12) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Apelin 12 (AP12) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Apelin 12 (AP12) 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)	90-97%	88-96%	87-97%	83-97%
EDTA plasma(n=5)	85-101%	97-105%	97-105%	97-104%
heparin plasma(n=5)	81-99%	91-102%	94-105%	98-105%

Stability

The stability of 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.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

爱帕琳肽12(AP12)检测试剂盒Assay procedure summary

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^oC;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37^oC;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 15-25 minutes at 37^oC;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

爱帕琳肽12(AP12)检测试剂盒Test principle

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Apelin 12 (AP12) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Apelin 12 (AP12) and unlabeled Apelin 12 (AP12) (Standards or samples) with the pre-coated antibody specific to Apelin 12 (AP12). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Apelin 12 (AP12) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Apelin 12 (AP12) in the sample.