Elisa Gluten Tests

Purpose

GlutenTox ELISA Sandwich   is   an  immunoabsorbent   test  for  the  detection  and quantification of the toxic fraction of gluten that is harmful to celiac disease sufferers.

Test Basis

GlutenTox ELISA Sandwich is a quantitative immunoabsorbent test designed for the detection and quantification of the toxic fraction of gluten. During the test, the sample is added to a plate coated with monoclonal anti-gliadin antibodies that specifically recognises the toxic fraction of gluten. After the washing steps, addition of a monoclonal anti-gliadin (G12)-HRP conjugated antibody will allow  to  measure  the  signal.

GlutenTox ELISA Sandwich is a quantitative method therefore  the higher concentration of gluten present the more intense the signal will be.

The ELISA  Sandwich  is a technique usually  used for the analysis of substances found at very low concentrations. This combined with the high specificity and sensitivity of the antibody used in this test allows this method to precisely quantify gluten in food samples.

Storage Conditions

•  Store all reagents of the kit at a temperature between 2 and 8 ºC. Do not freeze. Do not open until the moment of use. Reagents are stable up to the indicated expiration date if they are stored and handled correctly.
• Unused strips must be returned to the aluminum bag containing desiccants closed hermetically and stored between 2 to 8 ºC.
• The diluted Wash Solution is stable for two weeks at 2 to 8 ºC.

Precautions

• This kit is designed for professional in vitro use only.
• Do not use after expiration date.
• Do not mix the components of various kits or use reagents or solutions other than those provided.
• It is recommended to use non-powdered disposable gloves to work with this kit. Do not touch the strips with your hands.
• Incomplete sealing of the aluminum bag with the strips may cause accumulation of humidity and loss of precision in the results.
• It is recommended to strictly follow the instructions described in this manual.
• A number of factors influence the carrying out of the test (initial temperature of the reagents,  incubation time during the test, acuracy and reproducibility of the piping technique, quality of the washing technique). It is necessary to exercise extreme care and work in a consistent manner to obtain exact and reproducible results.
• The Stop Solution is composed of sulphuric acid (H2SO4). Avoid ingestion, inhalation, or contact with skin or eyes. Avoid exposure to bases, metals, or other compounds that may react with acids.
• The Substrate Solution is photosensitive; avoid prolonged exposure to light.

Supplied Materials

• 12 Microtiter strips (1×8, separable) coated with A1 anti-gliadin antibody. Ready to use.
• Extraction Solution (250 ml).
• Wash Solution 10x (40 ml).
• Dilution Solution (120 ml). Ready to use.
• Antibody GLUTENTOX G12-HRP conjugated (0.2 ml).
• Substrate Solution  (12 ml). Ready to use.
• STOP Solution, 1M H2SO4 (12 ml). Ready to use.
• Positive control (1.2 ml). Ready to use.
• Negative control  (1.2 ml). Ready to use.
• 5 GLUTENTOX standards (1.2 ml)  with 25; 12.5; 6.25; 3.12 and 1.56 ng/ml of gliadin. Ready to use.

Necessary materials not supplied

• Analytical laboratory scale
• Closing test tubes ( >10 ml)
• Test vials
• Micro-plate reader with 450 nm filters
• Disposable gloves
• Ethanol 60%
• Ethanol 80%
• Distilled water
• Timer
• Vortex agitator (optional)
• Laboratory tube rotators (optional)
• Centrifuge (optional)
• Automatic micro-plate washer (optional)
• Single channel pipettes, multichannel pipettes    (recommended) and tips