| Test principle: |
This assay employs a two-site sandwich ELISA to quantitate AQP4 in samples. An antibody specific for AQP4 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyAQP4 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for AQP4 is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of AQP4 bound in the initial step. The color development is stopped and the intensity of the color is measured.
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| Product Overview: |
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| Components: |
Reagents |
Quantity |
Reagents |
Quantity |
Assay plate (96 Wells) |
1 |
Instruction manual |
1 |
Standard (lyophilized) |
2 |
Sample Diluent |
1 x 20 mL |
Biotin-Conjugate (concentrate 100 x) |
1 x 120 μL |
Biotin-Conjugate Diluent |
1 x 20 mL |
Streptavidin-HRP (concentrate 100 x) |
1 x 120 μL |
Streptavidin-HRP Diluent |
1 x 20 mL |
Wash Buffer (concentrate 25 x) |
1 x 20 mL |
Substrate Solution |
1 x 12 mL |
Stop Solution |
1 x 10 mL |
Adhesive Films |
4 |
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| Specificity: |
This assay has high sensitivity and excellent specificity for detection of Human AQP4. No significant cross-reactivity or interference between Human AQP4 and analogues was observed.
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| Recovery: |
Matrices listed below were spiked with certain level of recombinant Human AQP4 and the recovery rates were calculated by comparing the measured value to the expected amount of Human AQP4 in samples.
Sample Type |
Number |
Recovery range (%) |
Average(%) |
Serum |
10 |
90-101 |
96 |
EDTA plasma |
10 |
89-97 |
93 |
Heparin plasma |
10 |
91-99 |
95 |
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| Precision: |
Intra-assay Precision (Precision within an assay)
Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-assay Precision (Precision between assays)
Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.
CV (%) = SD/meanX100
Intra-Assay: CV<8%
Inter-Assay: CV<12%
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| Linearity: |
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human AQP4 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample Type |
1：2 |
1：4 |
1：8 |
1：16 |
Serum |
78-89% |
81-99% |
92-103% |
95-105% |
EDTA plasma |
91-101% |
90-98% |
93-101% |
91-98% |
Heparin plasma |
92-103% |
93-102% |
92-99% |
91-101% |
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| Stability: |
The stability of ELISA 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.
The loss rate was determined by accelerated thermal degradation test. Keep the kit at 37°C for 4 and 7 days, and compare O.D.values of the kit kept at 37°C with that of at recommended temperature. (referring from China Biological Products Standard, which was calculated by the Arrhenius equation. For ELISA kit, 4 days storage at 37°C can be considered as 6 months at 2 - 8°C, which means 7 days at 37°C equaling 12 months at 2 - 8°C).
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| Sample collection and storage: |
S e r u m : U s e a s e r u m s e p a r a t o r t u b e ( S S T ) a n d a l l o w s a m p l e s t o c l o t f o r t w o h o u r s a t r o o m t e m p e r a t u r e o r o v e r n i g h t a t 2 - 8° C b e f o r e c e n t r i f u g a t i o n f o r 1 5 m i n u t e s a t 1 0 0 0 × g . R e m o v e s e r u m a n d a s s a y i m m e d i a t e l y o r a l i q u o t a n d s t o r e s a m p l e s a t ≤ - 2 0° C . A v o i d r e p e a t e d f r e e z e - t h a w c y c l e s .
P l a s m a : C o l l e c t p l a s m a u s i n g E D T A , o r h e p a r i n a s a n a n t i c o a g u l a n t . C e n t r i f u g e f o r 1 5 m i n u t e s a t 1 0 0 0 × g a t 2 - 8° C w i t h i n 3 0 m i n u t e s o f c o l l e c t i o n . A s s a y i m m e d i a t e l y o r a l i q u o t a n d s t o r e s a m p l e s a t ≤ - 2 0° C . A v o i d r e p e a t e d f r e e z e - t h a w c y c l e s .
O t h e r b i o l o g i c a l f l u i d s : C e n t r i f u g e s a m p l e s f o r 2 0 m i n u t e s a t 1 0 0 0 × g . R e m o v e p a r t i c u l a t e s a n d a s s a y i m m e d i a t e l y o r s t o r e s a m p l e s i n a l i q u o t a t - 2 0° C o r - 8 0° C . A v o i d r e p e a t e d f r e e z e / t h a w c y c l e s .
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| Kits storage instructions: |
Store at 2-8°C. Please refer to Instruction Manual.
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Human Aquaporin 4 (AQP4) ELISA Kit
