ELISA Kit for Apelin 13 (AP13)

APLN13

Specificity

This assay has high sensitivity and excellent specificity for detection of Apelin 13 (AP13).
No significant cross-reactivity or interference between Apelin 13 (AP13) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Apelin 13 (AP13) and the recovery rates were calculated by comparing the measured value to the expected amount of Apelin 13 (AP13) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 92-101 96
EDTA plasma(n=5) 92-101 96
heparin plasma(n=5) 90-99 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Apelin 13 (AP13) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Apelin 13 (AP13) 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 13 (AP13) 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) 80-101% 80-89% 95-102% 96-103%
EDTA plasma(n=5) 84-95% 87-101% 88-97% 80-89%
heparin plasma(n=5) 80-103% 83-91% 78-102% 92-105%

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

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
Investigative Ophthalmology and Visual Science Apelin in Plasma and Vitreous and in Fibrovascular Retinal Membranes of Patients with Proliferative Diabetic Retinopathy IOVS: 4237
Asian Pacific Journal of Cancer Prevention Is short-term exercise a therapeutic tool for improvement of cardioprotection against DOX-induced cardiotoxicity? An experimental controlled protocol in rats. Apjcpcontrol: Source
Clin Exp Hypertens Individual and concomitant effects of cardioprotective programs on cardiac apelinergic system and oxidative state in L-NAME-induced hypertension. PubMed: 22578103
Medical Science Monitor : International Medical Journal of Experimental and Clinical Research Effect of Ramadan fasting on serum concentration of apelin-13 and new obesity indices in healthy adult men NCBI: PMC3943714
ZJRMS Improvement of Kidney Apelin and Apelin Receptor in Nitro-L-Arginine-Methyl Ester Induced Rats Zjrms:Source
Iranian Journal of Kidney Diseases The Role of Apelin 13 in Progression of Chronic Kidney Disease. PubMed: 26338160
 Journal of Clinical & Experimental Nephrology Role of Circulating Serum Apelin-13 Levels in Glomerulonephritis: A Pilot Study aid:8469
international journal of cardiology Biochemical and genetic role of apelin in essential hypertension and acute coronarysyndrome. pubmed:27543713
Clinical and Experimental Hypertension Serum concentrations of apelin-17 isoform vary in accordance to blood pressure categories in individuals with obesity class 3 Pubmed:29652188
Progress in Nutrition Effect of fermented food consumption on biochemical parameters and adipokines levels Doi: 0.23751/pn.v20i4.7374
ACTA CARDIOLOGICA SINICA Impact of apelin-13 on the development of coronary artery ectasia Pubmed: 32425436
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