ELISA Kit for Apelin (APLN)

XNPEP2; APEL; APJ endogenous ligand

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 82-93 88
EDTA plasma(n=5) 94-101 98
heparin plasma(n=5) 83-98 89

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Apelin (APLN) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Apelin (APLN) 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) 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) 86-94% 81-97% 94-102% 94-105%
EDTA plasma(n=5) 95-103% 81-91% 87-102% 78-101%
heparin plasma(n=5) 99-105% 95-104% 78-96% 91-103%

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
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

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°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

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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
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