Multiplex Assay Kit for Adrenomedullin (ADM) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 92-105 97
EDTA plasma(n=5) 98-105 102
heparin plasma(n=5) 90-99 94

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Adrenomedullin (ADM) ,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 Adrenomedullin (ADM) ,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 Adrenomedullin (ADM) ,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) 81-95% 90-97% 92-105% 78-97%
EDTA plasma(n=5) 79-93% 80-101% 80-99% 83-101%
heparin plasma(n=5) 86-93% 99-105% 82-97% 78-101%

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#:ADM) 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
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
Biomedical Journal of Scientific & Technical Research Anti-Inflammatory and Pro-Inflammatory Neuropeptides in Dilative Cardiomyopathy are there Relations to Psychophysical Stress?
博士论文 Clinical and experimental investigation of the nervous system of neonatal foals. Neuroendocrin transition from foetal consciousness to perinatal life
Biology Hemodynamic Responses in Lower Limb Lymphedema Patients Undergoing Physical Therapy 34356497
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