Allergen ELISA Kit II: Crustacean

Article no: 1 500 016-2
  • Allergen detection in highly processed and heated food
  • High sensitivity (LOD: 0,31 ppm)
  • High specificity
  • Easy-to-use protocols
570,00 
678,30  incl. 19% VAT 
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Delivery status: Currently not in stock
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 1 500 016-2
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

Label Name of component Content Quantity
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 Crustacean Standard (50 ng/mL) 1.8 mL 1 vial
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.

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