Multiplex Assay Kit for Myelin Basic Protein (MBP) ,etc. by FLIA (Flow Luminescence Immunoassay)

Myelin A1 protein; Myelin membrane encephalitogenic protein

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 92-105 99
EDTA plasma(n=5) 91-105 99
heparin plasma(n=5) 81-89 86

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Myelin Basic Protein (MBP) ,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 Myelin Basic Protein (MBP) ,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 Myelin Basic Protein (MBP) ,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) 99-105% 95-105% 86-93% 80-105%
EDTA plasma(n=5) 82-94% 83-91% 97-105% 88-96%
heparin plasma(n=5) 92-101% 83-92% 96-104% 79-104%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:MBP) 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 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
PLOS ONE CD98 Positive Eosinophils Contribute to T Helper 1 Pattern Inflammation PlosOne: Source
Journal of Occupational Health Effects of Exposure to 1-Bromopropane on Astrocytes and Oligodendrocytes in Rat Brain Pubmed: 23183024
Pak J Med Sci Myelin basic protein and ischemia modified albumin levels in acute ischemic stroke cases PubMed: 26648996
Experimental and Toxicologic Pathology Morphology and staining behavior of neutrophilic and eosinophilic granulocytes of the common marmoset (Callithrix jacchus) Pubmed:27165445
Acta Angiologica Serum microtubule associated protein tau and myelin basic protein as the potential markers of brain ischaemia-reperfusion injury in patients undergoing carotid endarterectomy AA.2016.0008
Stem Cells. Myelin Basic Protein Regulates Primitive and Definitive Neural Stem Cell Proliferation from the Adult Spinal Cord. pubmed:27573615
Environmental Science and Pollution Research Glial markers and emotional memory in rats following acute cerebral radiofrequency exposures. pubmed:27696165
Acta Angiol Serum microtubule associated protein tau and myelin basic protein as the potential markers of brain ischaemia-reperfusion injury in patients undergoing carotid endarterectomy AA.2016.0008
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International Journal of Livestock Research Association and Risk of Canine Distemper with Respect to Age, Sex and Breed of Dogs Suffering from Demyelinating Neuropathies 10.5455:ijlr.20170811080455
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