These ELISA kits are sandwich enzyme immunoassays for the quantitative determination of individual proteins of allergenic ingredients in processed or unprocessed food. These kits are designed to be used by quality control personnel or other trained professionals, for allergen surveillance in the production or for product release controls.
Proteins in the samples are extracted, centrifuged and filtered prior to the sandwich ELISA. Antigen is bound to the polyclonal antibody coated wells of the microplate module resulting in an antigen-antibody complex. Subsequently, the enzyme-conjugated antibody is bound to the already bound antigen-antibody complex, forming an antibody-antigen-antibody sandwich.
Addition of enzyme substrate results in color development, and the intensity of which can be determined by the absorbance at 450 nm. The intensity of the color developed is directly proportional to the concentration of protein of allergenic ingredients in the food.
Scope of delivery
- 1 x 96 wells of microplates (12 strips with 8 wells)
- Associated buffers and enzymes
Specifications
| Article Number |
1 500 008-2 |
| Sample preparation time |
Approx. 10 min |
| Sample extraction time |
Approx. 30 min, or overnight |
| ELISA analysis |
Approx. 2 h |
| Assay Sensitivity |
0.31 ppm (μg protein /g food) |
| Detectable concentration range |
0.78 to 50 ppb detectable protein |
| Storage |
At 2-8°C (35-46°F), DO NOT FREEZE |
| Working temperature |
20-30°C (All solutions should be equilibrated) |
Kit components
| A |
Reagent A (10X Concentrate) |
100 mL |
1 Bottle |
| B |
Reagent B (10X Concentrate) |
100 mL |
1 Bottle |
| C |
Reagent C (10X Concentrate) |
100 mL |
1 Bottle |
| D |
Antibody-coated Microplate Module |
6X8 well modules |
2 packs |
| E |
Standard (50 ng/mL extracted protein solution) |
1 mL |
2 vials |
| F |
Enzyme-conjugated Antibody |
13 mL |
1 Bottle |
| G |
Enzyme Substrate (TMB Solution) |
13 mL |
1 Bottle |
| H |
Stop Solution (1N Sulfuric Acid) |
13 mL |
1 Bottle |
| I |
Wash Buffer (20X Concentrate) |
50 mL |
1 Bottle |
| |
Frame for mounting the microplate module |
|
1 Piece |
| |
Microplate cover |
|
1 Piece |
Instructions
Extraction procedure
Short time extraction method
- Grind/mince the samples
- Weigh 1.0 g sample in a tube, add 19 mL Sample Extraction Solution.
- Incubate capped tube in boiling water for 10 minutes.
- Cool down the tube and vortex for 30 sec.
- Adjust pH to 6.0-8.0.
- Centrifuge and filter the supernatant if necessary.
- Dilute Sample Extract 20-fold with Diluent I.
Overnight Extraction Method
- Grind/mince the samples
- Weigh 1.0 g sample in a tube, add 19 mL Sample Extraction Solution.
- Fix the tube to a shaker horizontally, and oscillate at room temperature overnight.
- Adjust pH to 6.0-8.0.
- Centrifuge and filter the supernatant if necessary.
- Dilute Sample Extract 20-fold with Diluent I.
ELISA procedure
- Pipette 100 μL Working Standard and Working Sample Solution into a well on the microplate.
- Incubate the microplate for 1 hour at 20-30°C (68-86°F).
- Wash the wells 6 times with Washing Solution.
- Dispense 100 μL Enzyme-conjugated Antibody (F).
- Incubate the microplate for 30 min at 20-30°C (68-86°F).
- Wash the wells 6 times.
- Dispense 100 μL Enzyme Substrate (G).
- Incubate reaction for 20 min at 20-30°C (68-86°F) in the dark.
- Stop the enzyme reaction by adding 100 μL Stop Solution (H).
- Measure absorbance at 450 nm.
- Calculate the concentration of detectable protein using standard curve.
