Multiplex Assay Kit for Apelin (APLN) ,etc. by FLIA (Flow Luminescence Immunoassay)

XNPEP2; APEL; APJ endogenous ligand

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Apelin (APLN) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Apelin (APLN) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Apelin (APLN) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Apelin (APLN) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 95-103 101
EDTA plasma(n=5) 95-102 99
heparin plasma(n=5) 86-93 89

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Apelin (APLN) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Apelin (APLN) ,etc. by FLIA (Flow Luminescence Immunoassay) 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 (APLN) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 94-101% 87-96% 91-98% 93-102%
EDTA plasma(n=5) 83-101% 82-104% 88-102% 80-95%
heparin plasma(n=5) 78-89% 97-105% 94-101% 80-96%

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

Reagents Quantity Reagents Quantity
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:APLN) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 50μL standard or sample to each well,
    add 10μL magnetic beads,and 50μL Detection Reagent A,incubate 60min at 37°C on shaker;
3. Wash plate on magnetic frame for three times;
4. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
5. Wash plate on magnetic frame for three times;
6. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Clin Exp Hypertens. Individual and Concomitant Effects of Cardioprotective Programs on Cardiac Apelinergic System and Oxidative State in └-NAME-Induced Hypertension Informahealthcare: Source
African Journal of Pharmacy and Pharmacology Cardioprotective effects of aerobic regular exercise against doxorubicin-induced oxidative stress in rat. Academicjournals: Source
Zahedan Journal of research in Medical Sciences Improvement of Kidney Apelin and Apelin Receptor in Nitro-L-Arginine-Methyl Ester Induced Hypertension Rats Zjrms: Source
Endocr J. CTRP3 modulates the expression and secretion of adipokines in 3T3-L1 adipocytes Pubmed:25168658
Mol Metab TIMP3 interplays with apelin to regulate cardiovascular metabolism in hypercholesterolemic mice PubMed: 26500845
Journal of medicinal food Regulation of GIP and Ghrelin in Healthy Subjects Fed on Sun-Dried Raisins: A Pilot Study with a Crossover Trial Design. pubmed:28170279
Arch Physiol Biochem Associations of apelin, leptin, irisin, ghrelin, insulin, glucose levels, and lipid parameters with physical activity during eight weeks of regular exercise training Pubmed: 31290696
preprints Serum Apelin and Galactin-3 in Preeclampsia in Iraq Pubmed: 32538210
Journal of Diabetes Research The Potential Role of Chemerin, Lipocalin 2, and Apelin in the Diagnosis and Pathophysiology of Gestational Diabetes Mellitus
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