OD Aflatoxins B1 Mycotoxin Elisa Test Kits For Feed Tissue Edible Oil 96 Wells/Kit

Aflatoxins B1 ELISA Test Kit for Feed Corn Peanut Tissue Edible oil 96 Wells/Kit

1. Aflatoxins B1 ELISA Test Kit Principle

This test kit is based on the indirect competitive enzyme immunoassay for the detection of Aflatoxin B1. The coupling antigen is pre-coated on the micro-well stripes. The Aflatoxin B1 in the sample and the coupling antigens pre-coated on the micro-well stripes compete for the anti- Aflatoxin B1 antibodies. After the addition of the enzyme conjugate, the TMB substrate is added for coloration. The optical density (OD) value of the sample has a negative correlation with the Aflatoxin B1 in the sample. This value is compared to the standard curve and the Aflatoxin B1 residues is subsequently obtained.

2. Technical specifications

Sensitivity: 0.02 ppb
Incubator temperature: 25℃
Incubator time: 30min～15min
Detection limit:
Feed, rice, corn, peanut, tissue, edible oil 2ppb
Cross-reaction rate:
Aflatoxin B1 100%
Aflatoxin B2 54.5%
Aflatoxin G1 8.4%
Aflatoxin G2 1.7%
Aflatoxin M1 6.6%
Recovery rate:
Feed, rice, corn, peanut, tissue, edible oil 95±35%

3. Aflatoxins B1 ELISA Test Kit Components

5. Sample pre-treatment

Instructions (The following points must be dealt with before the pre-treatment)
1) Only the disposable tips can be used for the experiments and the tips must be changed when used for absorbing different reagents;
2) Before the experiment, each experimental utensil must be clean and should be re-cleaned if necessary, in order to avoid the contamination that interferes with the experimental results.

Solution preparation before sample pre-treatment:
1) Sample redissolving solution
Use 1 part of 20X concentrated redissolving solution and dissolve with 19 parts of deionized water to obtain the ready to use sample redissolving solution.
2) Sample extract solution
Use 7 parts of Methanol and dissolve with 3 parts of deionized water to obtain the ready to use sample extract solution.

Samples Preparation
5.1 Preparation of tissue, feed, rice and corn sample
1) Take 1.0±0.05g grinded sample into 50ml centrifuge tube, add 5ml Sample extract solution, shake for 3min, centrifuge at above 4000r/min at 20℃ for 10 min;
2) Take 100ul supernatant, add 700ul Sample redissolving solution, shake to evenly;
3) Take 50μl to test
Dilution factor: 40

5.2 Preparation of edible oil sample
1) Take 1.0±0.05g edible oil sample into 50ml centrifuge tube; add 5ml sample extract solution, then add 4ml n-hexane, shake for 3min, centrifuge at above 4000r/min at 20℃ for 10 min;
2) Discard the supernatant, take 100ul of the middle-layer liquid, add 700ul Sample redissolving solution, shake to evenly;
3) Take 50μl to test
Dilution factor: 40

5.3 Preparation of peanut sample
1) Take 1.0±0.05g grinded peanut sample into 50ml centrifuge tube; add 5ml sample extract solution, then add 4ml n-hexane, shake for 3min, centrifuge at above 4000r/min at 20℃ for 10 min;
2) Discard the supernatant, take 100ul of the middle-layer liquid, add 400ul Sample redissolving solution, shake to evenly;
3) Take 50μl to test
Dilution factor: 25

6. ELISA procedures

6.1 Instructions
1) Bring ELISA reagents to room temperature (20 - 25 °C) before use.
2) Put ELISA reagents back to 2-8 ℃ immediately after use.
3) The ELISA reproducibility in the analysis process is largely depends on the consistency of the washing plate, the correct washing plate operation is the point of determination ELISA program.
4) In all process of constant temperature incubation, avoid light exposure, seal the microplate with the cover membrane.

6.2 Operation procedures
1) Bring test kit to the room temperature (20-25 ℃) for at least 30 min, note that each reagent must be shaken evenly before use;
2) Put the required micro-well strips into plate frames. Re-sealed the unused microplate, stored at 2-8 ℃, not frozen.
3) Solution preparation: take the 40ml 20× concentrated washing buffer, dissolve with deionized water at 1:19 (1 part 20× concentrated washing buffer + 19 parts deionized water), or prepare as quantity needed.
4) Numbering: number the micro-wells according to samples and standard solution; each sample and standard solution should be performed in duplicate; record their positions.
5) Add standard/sample: Add 50 µL of the sample or the standard solution into separate duplicate wells, then add enzyme conjugate, 50 µL/well; then antibody working solution, 50 µL/well. Mix gently by shaking the plate manually, seal the microplate with the cover membrane, incubate at 25 °C for 30 min in the dark.
6) Wash microplate: Carefully open the cover membrance, pour liquid out of microwell; add 250 µL/well of washing buffer, wash fully for 4-5 times, 15-30 s each time, then take out and flap to dry with absorbent paper.(Use unused spear to pierce bubble after dry)
7) Coloration: add 50 µL of substrate A solution then 50 µL substrate B solution into each well. Mix gently by shaking the plate manually, and incubate at 25 °C for 15min in the dark for coloration.
8) Determination: add 50 µL of the stop solution into each well. Mix gently by shaking the plate manually. Set the wavelength of the microplate reader at 450 nm to determine the OD value of every well. (Recommend to read the OD value at the dual-wavelength 450/630 nm within 5 min).

7. Precautions

1) The room temperature below 25 ℃ or the temperature of the reagents and the samples being not returned to the room temperature (20-25 ℃) will lead to a lower standard OD value.
2) Dryness of the microplate in the washing process will be accompanied by the situations including the non-linear standard curves and the undesirable reproducibility; So continue to next step immediately after washing.
3) Mix evenly before adding any reagents.
4) The stop solution is the 2 M sulfuric acid solution, avoid contacting with the skin.
5) Do not use the kit exceeding its expiry date. The use of diluted or adulterated reagents from the kits will lead to the changes in the sensitivity and the detecting OD values. Do not exchange the reagents from the kits of different lot numbers to use.
6) Storage: store at 2-8 ℃, not frozen. Put the unused microplate into an auto-sealing bag to re-seal it. The standard substance and the colourless color former is light sensitive, and thus they cannot be directly exposed to the light.
7) Discard the colouration solution with any color that indicates the degeneration of this solution. The detecting value(450/630nm) of the 0 standard solution (0 ppb) of less than 0.5((A450nm<0.5)) indicates its degeneration.
8) The optimum reaction temperature is 25 ℃, and too high or too low temperatures will result in the changes in the detecting sensitivity and OD values.

8. Storage and expiry date

Storage: store at 2-8 ℃, not frozen.
Expiry date: 12 months; date of production is on box.
Note: If the Vacuum package of microplate has leakage, it is still valid to use, do not affect the test result, be relax to use.

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.