Multiplex Assay Kit for Aquaporin 4 (AQP4) ,etc. by CBA (Cytometric Bead Array)

HMIWC2; MIWC; WCH4; Mercurial-insensitive water channel

Specificity

This assay has high sensitivity and excellent specificity for detection of Aquaporin 4 (AQP4) ,etc. by CBA (Cytometric Bead Array).
No significant cross-reactivity or interference between Aquaporin 4 (AQP4) ,etc. by CBA (Cytometric Bead Array) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Aquaporin 4 (AQP4) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Aquaporin 4 (AQP4) ,etc. by CBA (Cytometric Bead Array) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 80-99 81
EDTA plasma(n=5) 91-103 96
heparin plasma(n=5) 93-101 96

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Aquaporin 4 (AQP4) ,etc. by CBA (Cytometric Bead Array) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Aquaporin 4 (AQP4) ,etc. by CBA (Cytometric Bead Array) 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 Aquaporin 4 (AQP4) ,etc. by CBA (Cytometric Bead Array) 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) 94-101% 81-93% 92-102% 98-105%
EDTA plasma(n=5) 80-95% 91-98% 93-101% 84-99%
heparin plasma(n=5) 80-96% 79-99% 90-99% 95-103%

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

Assay procedure summary

1. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 100μL standard or sample to each well, add 10μL magnetic beads, and incubate 120 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60 minutes protect from light at 25°C on shaker;
5. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
6. Wash plate once on magnetic frame;
7. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.

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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
Basic and Clinical Neuroscience Paeonol Protects Against Intrastriatal 6-Hydroxydopamine Rat Model of Parkinson's Disease 33995926
PLoS One Evaluation of aquaporins in the cerebrospinal fluid in patients with idiopathic normal pressure hydrocephalus 34597351
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