Intended Use
Enzyme Immunoassay for the Quantitative Determination of Gliadin/Gluten in Food
Contents of Kit
The kit contains reagents for 96 determinations. They have to be stored at 2-8°C. Expiry data are found on the labels of the bottles and the outer package.
1. Microtiter plate consisting of 12 strips with 8 breakable wells each, coated with anti-gliadin.
2. Gliadin Standards (0, 2, 6, 20, 60 ppm Gliadin): 1 × 5 vials with 1.0 mL each, dyed red, ready-touse.
3. Conjugate (anti-gliadin-Peroxidase): 15 mL, dyed red, ready-to-use.
4. Substrate Solution (TMB): 15 mL, ready-to-use.
5. Stop Solution (0.5 M H2SO4): 15 mL, ready-touse.
6. Sample dilution buffer (Tris): 1 × 60 mL as 10x concentrate, dyed red. Dilute 1+9 with distilled water. Stored at 4°C the diluted buffer is stable for at least one week. If during the cold storage crystals precipitate, the concentrate should be warmed up to 37°C for 15 minutes.
7. Washing Solution (PBS + Tween 20): 60 mL as 10× concentrate, dyed blue. Dilute 1+9 with distilled water. If during the cold storage crystals precipitate, the concentrate should be warmed up to 37°C for 15 minutes.
8. Two plastic foils to cover the strips during the incubation.
9. Plastic bag to store unused microtiter strips.
10. Instruction Manual.
Storage
Stored at 2-8°C. Expiry data are found on the labels of the bottles and the outer package. For more detailed information, please download the following document on our website.
Precision
Intra-assay Precision: 4 - 5%
Inter-assay Precision: 2 - 3%
Inter-lot Precision: 5 - 13%
Sensitivity
The limit of detection (LOD) of the Gliadin/ Gluten test is 0.3 ppm Gliadin. The limit of quantification (LOQ) of the Gliadin/Gluten test is 2 ppm Gliadin. Due to the variety of sample matrices and their influence on the blank, results less than the LOQ should be treated as negative.
General Description
Gluten is the main part of the protein fraction of cereals and consists of nearly the equal amount of the protein compounds prolamin (gliadin) and glutenin. Because of its special physico-chemical attributes and its low price, gluten is not only contained in cereal products, but also in other food as sausage products and ice cream or in drugs and cosmetics as binder and filler. For some persons, gluten has a pathological effect (coeliac disease). These people need to have a strict gluten free diet. In the European Union a maximum level of 20 ppm gluten is allowed for products declared as "gluten-free", and 100 ppm gluten for products declared as "very low gluten" respectively. Sensitive detection systems are required to determine gluten residues in foodstuff. The Gliadin/Gluten ELISA represents a highly sensitive detection system and is particularly capable of the quantification of gliadin/gluten residues in bakery products, baby food, meat and chocolate.
Citations
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Gliadin is a class of proteins found in wheat and several other cereals belonging to the grass genus Triticum. It is a crucial component of gluten, the protein complex responsible for the elasticity and rising properties of bread during baking. Gliadins, along with glutenins, constitute the main constituents of the gluten fraction present in wheat seeds and products such as wheat flour. There are three primary types of gliadin: α, γ, and ω, which are distinguished based on their amino acid sequences in the N-terminal cysteine domain.
Gliadins are intrinsically disordered proteins, meaning they have continuously altering shapes, making them difficult to study. However, research suggests they likely have a tadpole-like structure with a hydrophobic core and a loose, disordered tail. Unlike glutenins, which form extended networks of polymers due to disulfide bonds, gliadins are monomeric molecules in the cell, with their cysteines forming intra-chain disulfide bonds during synthesis. Gliadins are capable of aggregating into larger oligomers and interacting with other gluten proteins due to their large hydrophobic sections, poly-glutamine, and repetitive sequences. These properties contribute to their ability to form gluten and their role in food-derived pathogenesis.
Individuals with celiac disease, a chronic, immune-mediated intestinal disorder, exhibit a lifelong intolerance to gliadins and other prolamins found in wheat, barley, and rye. The immune system of these individuals recognizes specific amino acid sequences within the gliadin proteins, triggering an abnormal immune response and intestinal damage. Deamidated gliadin, produced by enzymatic or acid treatment, is even more reactive and can lead to more severe symptoms in those with celiac disease.
Alternative Names
Gliadin ELISA
Gluten ELISA
Gluten ELISA Kit
Gliadin cytotoxicity and in vitro cell cultures
Toxicology Letters
Authors: Elli L, Dolfini E, Bardella M T.
Abstract
Interest in celiac disease, a common enteropathy in Europe and the USA (1/200) caused by the dietary ingestion of gluten in susceptible subjects, has increased over the last few years. Its pathogenesis is still not completely clear, but it certainly involves immune-mediated mechanisms. Although a number of studies have been published concerning the role of T cells in inducing intestinal damage, little is known about the early stages in which gliadin (the toxic component of gluten) starts the whole process. In vitro two- and three-dimensional (multicellular spheroid) cell cultures are a simple and useful means of studying the direct cellular effects of gliadin and other "toxic" cereal peptides. Furthermore, in addition to improving our understanding of pathogenetic mechanisms, cell cultures can also be used to test modified peptides that could replace the toxic components present in the foods that celiac patients must avoid.
Relation between gliadin structure and coeliac toxicity
Acta Paediatrica
Authors: Wieser H.
Abstract
Gliadin, the alcohol-soluble protein fraction of wheat, contains the factor toxic for coeliac patients. The numerous components of gliadin can be classified according to their primary structure into ω5-, ω1,2-, α- and γ-type. Both ω- types have almost entirely repetitive amino acid sequences consisting of glut amine, proline and phenylalanine, α-and γ-type gliadins contain four and five different domains, respectively, and are homologous within the domains III and V. Unique for each α-and γ-type is domain I, which consists mostly of repetitive sequences rich in glutamine, proline and aromatic amino acids. Coeliac toxicity of gliadin is not destroyed by digestion with gastropancreatic enzymes. In vivo testing established the toxicity of α-type gliadins and in vitro testing of gliadin peptides revealed that domain I of α-type gliadins is involved in activating coeliac disease. The sequences -Pro-Ser-Gln-Gln- and -Gln-Gln-Gln-Pro- were demonstrated to be common for toxic gliadin peptides. Most of the in vivo and in vitro studies of synthetic peptides confirmed the importance of one or both of these sequences. Cultivated hexaploid, tetraploid and diploid wheat species do not differ significantly in potential toxic sequences of α-type gliadins.
COA
Certificate of Analysis
