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Nitrofuran AHD Diagnostic ELISA Kit for Aquatic Products 96Wells/Kit
1. Nitrofuran AHD Diagnostic ELISA Kit Principle
This kit is based on a competitive enzyme immunoassay for the
detection of aminohydantoin (AHD) in samples. Conjugated antigens
are pre-coated on microwell strips. Aminooxalic acid (AHD) in the
sample competes with the conjugated antigen pre-coated on the
microwell strip for anti-AHD antibodies. After the enzyme conjugate
is added, TMB substrate is added for staining. The optical density
(OD) value of a sample is negatively correlated with the AHD in it.
This value is compared to a standard curve and the AHD
concentration is subsequently obtained.
2. Technical Specifications
Sensitivity: 0.04ppb
Incubation temperature: 25℃
Incubation time: 30min~15min
Detection limit tissue, eggs, honey: 0.1ppb
cross-reaction rate
AHD 100%
AMOZ < 0.1%
AOZ < 0.1%
SEM < 0.1%
recovery rate
Tissue, eggs 95±25%
Honey 75±25%
3. Nitrofuran AHD Diagnostic ELISA Kit Components
1 | Micro-well strips | 12 strips with 8 removable wells each | |
2 | 6× standard solution (1 mL each) | 0ppb | 0.04ppb |
0.12ppb | 0.36ppb | ||
1.08ppb | 3.24ppb | ||
3 | Enzyme conjugate | 7ml | red cap |
4 | Antibody working solution | 7ml | blue cap |
5 | Substrate A | 7ml | white cap |
6 | SubstrateB | 7ml | black cap |
7 | Stop solution | 7ml | yellow cap |
8 | 20× concentrated washing buffer | 40ml | white cap |
9 | 2× concentrated redissolving solution | 50ml | transparent cap |
10 | 2-Nitrobenzaldehyde (C7H5NO3) | 10ml | black cap |
4. Sample preparation
Tissue, eggs
1) Weigh 1±0.05g of homogeneous sample, add 4mL of distilled water,
0.5mL of 1 M HCl and 100µL of 2-nitrobenzaldehyde (C7H5NO3) to each
tube, and shake appropriately for 2min;
2) Incubate overnight at 37°C (about 16 hours) or in a water bath
at 56°C (2 hours).
3) Add 5mL 0.1M K2HPO4, 0.4mL 1M NaOH and 6mL ethyl acetate to each
tube, shake well for 30s.
4) Centrifuge at room temperature (20-25°C) above 4000r/min for
10min (if there is emulsification or the ethyl acetate layer is
less than 3ml, incubate the sample in a water bath at 80°C for
10min, and centrifuge repeatedly; or increase the speed and prolong
the centrifugation time).
5) Transfer 3 mL of ethyl acetate layer to a new centrifuge tube
and evaporate to dryness with nitrogen or air at 50°C.
6) Dissolve the dry residue with 2 mL of n-hexane, add 1 mL of the
diluted reconstituted solution, mix well for 30 seconds, and
centrifuge at room temperature (20-25°C) at 4000 rpm for 10
minutes; remove the upper n-hexane phase (if emulsified)
phenomenon, after removing the upper n-hexane phase, the samples
were incubated in a 70°C water bath for 10-20min, and centrifuged
repeatedly).
7) Take 50 µL of the lower layer for analysis.
Sample dilution factor: 2
5. matters needing attention
1) If the room temperature is lower than 25°C or the reagent
temperature and the sample do not return to room temperature
(20-25°C), the standard OD value will be low.
2) During the washing process, the drying of the microplate will be
accompanied by the nonlinearity of the standard curve, and the
reproducibility is not ideal; therefore, proceed to the next step
immediately after washing.
3) Mix well, otherwise the reproducibility will be poor.
4) The stop solution is 2M sulfuric acid solution, avoid contact
with skin.
5) Do not use the kit beyond the shelf life. The use of diluted or
adulterated reagents from the kit can result in changes in
sensitivity and detection OD values. Do not replace reagents in
different batches of kits.
6) Reseal unused microplates in ziplock bags. Standard solutions
and colorless couplers are light-sensitive and cannot be directly
exposed to light.
7) Discard any tinting solution whose color indicates that the
solution has been denatured. A detection value of less than 0.5 for
Standard Solution 1 (0 ppb) indicates degradation.
8) The optimal reaction temperature is 25°C, and the detection
sensitivity and OD value will change if the temperature is too high
or too low.
6. Storage and validity period
Storage: Store at 2 to 8°C, do not freeze.
Expiration date: 12 months; production date is on the box.
Note: If the microplate vacuum packaging leaks, it can still be
used without affecting the test results, please use it with
confidence.
Q1: When will it be shipped?
A1: We will ship the goods for you as soon as possible within 7
working days after receiving the payment. (In case of external
factors such as the epidemic, the delivery may be delayed)
Q2: Does it support OEM/ODM?
A2: It can be supported, but the specific quantity needs to be more
than 100,000 pieces, which is convenient for customized products.
Q3: How is your factory doing in terms of quality control?
A3: We have nationally certified ISO9001 and ISO13485. Our
production process conforms to standard procedures to ensure
optimum product quality.
Q4: How to provide after-sales service?
A4: We provide professional online technical after-sales service.
We can provide you with one-on-one guidance via video, telephone,
etc.
Q5: What is the payment method?
A5: We receive payment by T/T.
Q6: How to ship?
A6: Choose the best shipping method for you by getting quotes from
our many cooperative carriers, and also ship according to your
requirements.