ELISA Kit for Anti-Myelin Basic Protein Antibody (Anti-MBP)

Myelin A1 protein; Myelin membrane encephalitogenic protein

Specificity

This assay has high sensitivity and excellent specificity for detection of Anti-Myelin Basic Protein Antibody (Anti-MBP).
No significant cross-reactivity or interference between Anti-Myelin Basic Protein Antibody (Anti-MBP) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of Anti-Myelin Basic Protein Antibody (Anti-MBP) and the recovery rates were calculated by comparing the measured value to the expected amount of Anti-Myelin Basic Protein Antibody (Anti-MBP) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 80-101 91
EDTA plasma(n=5) 81-99 94
heparin plasma(n=5) 94-102 98

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Anti-Myelin Basic Protein Antibody (Anti-MBP) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Anti-Myelin Basic Protein Antibody (Anti-MBP) 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 Anti-Myelin Basic Protein Antibody (Anti-MBP) 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% 88-98% 87-97% 98-105%
EDTA plasma(n=5) 99-105% 91-102% 80-101% 87-101%
heparin plasma(n=5) 82-89% 95-102% 80-104% 92-101%

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
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
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 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 5 times;
5. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
6. Add 50µL Stop Solution. Read at 450nm immediately.

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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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