ELISA Kit for Anti-Aquaporin 4 Antibody (Anti-AQP4)

HMIWC2; MIWC; WCH4; Mercurial-insensitive water channel

Specificity

This assay has high sensitivity and excellent specificity for detection of Anti-Aquaporin 4 Antibody (Anti-AQP4).
No significant cross-reactivity or interference between Anti-Aquaporin 4 Antibody (Anti-AQP4) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of Anti-Aquaporin 4 Antibody (Anti-AQP4) and the recovery rates were calculated by comparing the measured value to the expected amount of Anti-Aquaporin 4 Antibody (Anti-AQP4) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 97-104 101
EDTA plasma(n=5) 93-101 97
heparin plasma(n=5) 99-105 102

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Anti-Aquaporin 4 Antibody (Anti-AQP4) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Anti-Aquaporin 4 Antibody (Anti-AQP4) 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-Aquaporin 4 Antibody (Anti-AQP4) 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) 89-103% 98-105% 78-90% 82-89%
EDTA plasma(n=5) 89-97% 93-101% 84-98% 96-105%
heparin plasma(n=5) 88-103% 78-98% 78-102% 96-103%

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
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
Neuroscience Letters Aquaporins AQP1 and AQP4 in the cerebrospinal fluid of bacterial meningitis patients ScienceDirect: S0304394011012341
Neurology Astrocytic damage is far more severe than demyelination in NMO Neurology: 208
Fluids and Barriers of the CNS Aquaporin-4 expression in the cerebrospinal fluid in congenital human hydrocephalus PubMed: PMC3651869
Neuropeptides Melanocortin MC4 receptor agonists alleviate brain damage in abdominal compartment syndrome in the rat PubMed: 25616531
Human & Experimental Toxicology Novel effect of Daflon and low-dose γ-radiation in modulation of thioacetamide-induced hepatic encephalopathy in male albino rats Pubmed:26987350
Cephalalgia Decreased levels of aquaporin-4 in the cerebrospinal fluid of patients with idiopathic intracranial hypertension Pubmed:26853804
PLoS One Cerebrospinal fluid biomarkers of infantile congenital hydrocephalus. pubmed:28212403
Journal of Neurotrauma Neuron-derived Plasma Exosome Proteins after Remote Traumatic Brain Injury Pubmed: 31441374
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