AF(Total Aflatoxin) ELISA Kit

Rs. 44,175.00
SKU E-TO-E006

Catalog No: E-TO-E006

Test principle:

This kit uses Indirect-Competitive-ELISA as the method. It can detect Aflatoxin (AF) in samples, such as grain, peanut, formula feed, etc. This kit is composed of ELISA Microtiter plate, HRP conjugate, antibody, standard, and other supplementary reagents. The microplate provided in this kit has been pre-coated with AF. During the reaction, AF in the samples or standard competes with AF on the solid phase supporter for sites of AF antibody. Then Horseradish Peroxidase (HRP) conjugate is added to each microplate well, and TMB substrate is for color development. There is a negative correlation between the OD value of samples and the concentration of AF. The concentration of AF in the samples can be calculated by comparing the OD of the samples to the standard curve.

Technical indicator:

Sensitivity: 0.02 ppb (ng/mL)

Reaction mode: 25℃, 30 min~ 15 min

Detection limit: 

Grain---0.1 ppb; Formula feed---0.2 ppb; Edible oil/Peanut---0.2 ppb;

Sauce/Wheat/Barley feed---0.2 ppb; beer---0.2 ppb; Wine/Soy sauce/Vinegar---0.1 ppb

Cross-reactivity: 

Aflatoxin B1 (AFB1) ---100%, AflatoxinB2 (AFB2) ---80%,

AflatoxinG1 (AFG1)--75%, AflatoxinG2 (AFG2) ---45%, FlatoxinM1 (AFM1) ---8%

Sample recovery rate: 

Grain/Formula feed---85%±15%, Peanut---82%±15%, Edible oil---85%±15%

Sauce/Wheat/Barley feed---83%±15%, Beer---84%±15%,

Wine/Soy sauce/Vinegar---87%±15%

 Kits components

Item

Specifications

ELISA Microtiter plate

96 wells

Standard Liquid

1 mL each

(0 ppb, 0.02 ppb, 0.04 ppb, 0.08 ppb, 0.16 ppb, 0.32 ppb)

High Concentrated Standard (100ppb)

1 mL

HRP Conjugate

5.5 mL

Antibody Working Solution

5.5 mL

Substrate Reagent A

6 mL

Substrate Reagent B

6 mL

Stop Solution

6 mL

20×Concentrated Wash Buffer

40 mL

Plate Sealer

3 pieces

Sealed Bag

1 piece

Manual

1 copy

 

Other supplies required

Instrument: Microplate reader, Printer, Homogenizer, Nitrogen Evaporators, Oscillators, Centrifuge, Graduated pipette, Balance (sensibility 0.01 g).

High-precision transferpettor: Single channel (20-200 μL, 100-1000 μL), Multichannel (300 μL).

Reagents: Methanol, N-hexane, Chloroform or Dichloromethane.

Assay procedure

Centrifuge the sample again after thawing before the assay. Bring all reagents to room temperature for 30 min before use. All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. 

  1. Number: number the sample and standard in order (multiple wells), and keep a record of standard wells and sample wells.
  2. Add sample: add 50 μL of Standard or Sample per well, then add 50 μL of HRP conjugate to each well, then add 50 μL of antibody working solution, cover the plate sealer oscillate for 5 s gently to mix thoroughly, incubate for 30 min at 25.
  3. Wash: uncover the sealer carefully, remove the liquid in each well. Immediately add 300 μL of wash working buffer to each well and wash. Repeat wash procedure for 5 times, 30 s intervals/time. Invert the plate and pat it against thick clean absorbent paper (If bubbles exist in the wells, clean tips can be used to prick them).
  4. Color Development: add 50 μL of substrate solution A to each well, and then add 50 μL of substrate solution B. Gently oscillate for 5 s to mix thoroughly. Incubate shading light for 15 min at 25.
  5. Stop reaction: add 50 μL of stop solution to each well, oscillate gently to mix thoroughly.
  6. OD Measurement: determine the optical density (OD value) of each well at 450 nm with a microplate reader (the 450/630 nm double wavelength is recommended). This step should be finished in 10 min after stop reaction.

Storage and valid period

Storage: Store at 2-8℃. Avoid freeze/thaw cycles.

Valid Period: 1 year, the expiration date is on the packing box.

 

Download Manual

 

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