ELISA Kit for Adrenomedullin (ADM)

AM; PAMP; Proadrenomedullin N-20 terminal peptide; ProAM N-terminal 20 peptide

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 88-95 92
EDTA plasma(n=5) 79-96 91
heparin plasma(n=5) 78-103 84

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Adrenomedullin (ADM) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Adrenomedullin (ADM) 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 Adrenomedullin (ADM) 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) 93-102% 82-101% 96-104% 93-102%
EDTA plasma(n=5) 89-98% 80-99% 85-93% 78-103%
heparin plasma(n=5) 96-103% 85-102% 95-102% 91-104%

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 Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
Reagent Diluent 1×300µL Stop Solution 1×6mL
TMB Substrate 1×9mL Instruction manual 1
Wash Buffer (30 × concentrate) 1×20mL

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
Journal of Periodontal Research Gingival crevicular fluid adrenomedullin level in individuals with and without diabetes mellitus type 2 Wiley: Source
J Periodontal Res. Comparison of peri-implant crevicular fluid levels of adrenomedullin and human beta defensins 1 and 2 from mandibular implants with different implant stability quotient levels in nonsmoker patients Pubmed: 23898869
Journal of Biosciences Short-term hypoxia/reoxygenation activates the angiogenic pathway in rat caudate putamen. Pubmed: 23660671
Journal of Translational Medicine DNA methylation patterns in newborns exposed to tobacco in utero Pubmed:25623364
Pediatric Hematology and Oncology Adrenomedullin—A New Marker in Febrile Neutropenia: Comparison With CRP and Procalcitonin PubMed: 26271020
Biogerontology. Angiogenesis is VEGF-independent in the aged striatum of male rats exposed to acute hypoxia. pubmed:28501895
Journal of Clinical Investigation  Adrenomedullin and Galanin Responses to Orthostasis in Older Persons pubmed:28796366
Clinical Pathology and Medical Laboratory ADRENOMEDULIN DI KARSINOMA PAYUDARA DENGAN METASTASIS :
Phytomedicine Composition and cardioprotective effects of Primula veris L. solid herbal extract in experimental chronic heart failure Doi: 10.1016/j.phymed.2018.09.015
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