ELISA Kit for Neurogranin (NRGN)

Ng; NEUG; Protein Kinase C Substrate RC3

Specificity

This assay has high sensitivity for detection of Neurogranin (NRGN).
100% cross-reactivity of Neurogranin (NRGN) was observed among Human, Mouse, Rat.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 89-104 93
EDTA plasma(n=5) 91-98 94
heparin plasma(n=5) 84-102 90

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Neurogranin (NRGN) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Neurogranin (NRGN) 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 Neurogranin (NRGN) 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) 85-103% 98-105% 91-105% 90-104%
EDTA plasma(n=5) 87-103% 95-103% 95-104% 86-97%
heparin plasma(n=5) 78-94% 82-99% 81-89% 80-103%

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
Alzheimer's & Dementia: Diagnosis, Assessment & Disease Monitoring Prediction of conversion from mild cognitive impairment to dementia with neuronally derived blood exosome protein profile Pubmed:27408937
Annals of Clinical and Translational Neurology Plasma neuronal exosomal levels of Alzheimer's disease biomarkers in normal aging Pubmed:27231710
The FASEB Journal Decreased synaptic proteins in neuronal exosomes of frontotemporal dementia and Alzheimer’s disease pubmed:27601437
Journal of Alzheimer's Disease Growth Hormone-Releasing Hormone Administration in Mild Cognitive Impairment and Its Impact on Neuronal Exosome Biomarkers of Dementia Doi: 10.3233/JAD-180302
SYNAPTIC PROTEIN BIOMARKERS AND DIFFERENTIAL DIAGNOSIS OF ALZHEIMER'S DISEASE AND OTHER NEURODEGENERATIVE DISORDERS
Am J Emerg Med The diagnostic value of neurogranin in patients with carbon monoxide poisoning: Can it show early neurological damage? 34388687
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