Wheat Gliadin IgG ELISA Kit

Name: Wheat Gliadin IgG ELISA Kit
SKU: DEIA274
Rating: 5 (1 reviews)

Regulatory status: For research use only, not for use in diagnostic procedures.

COA

SDS

Datasheet

Summary

Sample

Serum and plasma

Species Reactivity

Human

Detection Method

iELISA

Intended Use

The Gliadin IgG Antibody ELISA Test Kit has been designed for the detection and the quantitative determination of specific IgG antibodies against Gliadin in serum and plasma. Further applications in other body fluids are possible and can be requested from the Technical Service. This assay is intended for research use only.

Contents of Kit

Store kit components at 2-8°C and do not use after the expiry date on the box outer label. Before use, all components should be allowed to warm up to ambient temperature (18- 25°C). After use, the plate should be resealed, the bottle caps replaced and tightened and the kit stored at 2-8°C. The opened kit should be used within three months.
Mikrotiter Strips: 12 strips with 8 breakable wells each, coated with a Gliadin antigen (purified gluten antigen from wheat). Ready-to-use.
Calibrator A (Negative Control): 2 mL, protein solution diluted with PBS, contains no IgG antibodies against Gliadin. Addition of 0.01 % methylisothiazolone and 0.01 % bromonitrodioxane. Ready-to-use.
Calibrator B (Cut-Off Standard): 2 mL human serum diluted with PBS, contains a low concentration of IgG antibodies against Gliadin. Addition of 0.01 % methylisothiazolone and 0.01 % bromonitrodioxane. Ready-to-use.
Calibrator C (Weak Positive Control): 2 mL, human serum diluted with PBS, contains a medium concentration of IgG antibodies against Gliadin. Addition of 0.01 % methylisothiazolone and 0.01% bromonitrodioxane. Ready-to-use.
Calibrator D (Positive Control): 2 mL, human serum dilute with PBS, contains a high concentration of IgG antibodies against Gliadin. Addition of 0.01 % methylisothiazolone and 0.01 % bromonitrodioxane. Ready-to-use.
Enzyme Conjugate: 15 mL, anti-human-IgG-HRP (rabbit), in protein-containing buffer solution. Ready-to-use.
Substrate: 15 mL, TMB (tetramethylbenzidine). Ready-touse.
Stop Solution: 15 mL, 0.5 M sulfuric acid. Ready-to-use.
Sample Diluent: 60 mL, PBS/BSA buffer. Addition of 0.095% sodium azide. Ready-to-use.
Washing Buffer: 60 mL, PBS+Tween 20, 10× concentrate. Final concentration: dilute 1+9 with distilled water. If during the cold storage crystals precipitate, the concentrate should be warmed up at 37°C for 15 minutes.
Plastic Foils: 2 pieces to cover the microtiter strips during the incubation.
Plastic Bag: Resealable, for the dry storage of non-used strips.

Storage

Store kit at 2-8°C. It is stable up to the expiry date stated on the label of the box. Do not use kit beyond its expiry date.

Performance Characteristics

General Description

Gliadin is the main component of gluten, which occurs in wheat and other domestic grain types like rye, barley and oats, and may lead to severe diseases of the intestinal mucosa in sensitive children and adults. Celiac disease, a gluten-induced enteropathy, appears rather frequently (1 case on 300 births) and is a typical example of a non-IgE mediated food allergy. Genetically, histocompatibility antigens on the chromosome 6 are responsible for the disease. Celiac disease manifests itself practically as a constant reaction against gliadin. By the toxic effect of gluten in the intestinal tract, antibodies, cytokines and lymphocytes are released, which lead to internal lesions and inflammations. Further, the microvilli of the intestine are almost completely reduced, so that the inner intestinal surface becomes flat. The resulting malabsorption leads to a deficit of above all trace elements and vitamins. Loss of weight, diarrhea, flatulence and abdominal pain are observed as symptoms. An invasive diagnostic possibility represents the biopsy of the intestinal mucosa. In addition serological methods for the determination of IgG and IgA antibodies against gliadin, reticulin and endomysium in the patient serum are increasingly used as a screening method. For children with a gluten-sensitive enteropathy, the incidence was calculated to 90-100%, for adults with celiac disease 75- 90% and for dermatitis herpetiformis 40-50%. Elevated levels of IgA anti-gliadin demonstrate an active process and are in close correlation with a villous atrophy in children. The ELISA antibody determination is also well suited for the monitoring of patients after a gluten-free diet.

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Documents

pdf COA

pdf SDS

pdf Datasheet

QA & Reviews

Customer Reviews

References

Understanding the role of gluten subunits (LMW, HMW glutenins and gliadin) in the networking behavior of a weak soft wheat dough and a strong semolina wheat flour dough and the relationship with linear and non-linear rheology

FOOD HYDROCOLLOIDS

Authors: Bonilla, Jose C.; Erturk, Merve Y.; Kokini, Jozef L.

Abstract

The differences in viscoelastic properties of gluten from two very different wheat flours, a weak soft flour dough and a strong semolina dough, primarily caused by their gliadin and glutenin content including gliadin to glutenin ratio were studied. The doughs were subjected to oscillatory time sweep tests at small and large amplitudes as well as small and large frequencies. The gluten subunits (LMW, HMW glutenins and gliadins) tagged with specific fluorescent quantum dots with a distint excitation wavelength were imaged with confocal laser scanning microscopy and their distribution and interactions were measured. The quantitative imaging data was converted into networking data that included lacunarity, network area, and number of network junctions. This networking data was correlated with rheology. The two different dough systems showed different oscillatory behavior during time sweeps. The critical role of LMW glutenins in keeping the structural integrity of semolina doughs was demonstrated by a direct correlation between the non-linear elastic component 'e(3)/e(1)' of the dough and protein network parameters of LMW glutenins. It was also shown that gliadins and HMW glutenins co-localize and associate throughout rheological deformations of the dough. The disruption of the three gluten subunits are responsible for rheological breakdown of soft wheat flour dough, with gliadins influencing the breakdown of the network at higher amplitude deformations. This research presents a new method to analyze the microstructure of wheat doughs and new understanding of how the network structure of the dough subunits contribute and are correlated with fundamental rheological tests.

Identification of Isopeptides Between Human Tissue Transglutaminase and Wheat, Rye, and Barley Gluten Peptides

SCIENTIFIC REPORTS

Authors: Lexhaller, Barbara; Ludwig, Christina; Scherf, Katharina Anne

Abstract

Celiac disease (CD) is a chronic immune-mediated enteropathy of the small intestine, which is triggered by the ingestion of storage proteins (gluten) from wheat, rye, and barley in genetically predisposed individuals. Human tissue transglutaminase (TG2) plays a central role in the pathogenesis of CD, because it is responsible for specific gluten peptide deamidation and covalent crosslinking, resulting in the formation of N-epsilon-(gamma-glutamyl)-lysine isopeptide bonds. The resulting TG2-gluten peptide complexes are assumed to cause the secretion of anti-TG2 autoantibodies, but the underlying mechanisms are only partly known. To gain more insight into the structures of these complexes, the aim of our study was to identify TG2-gluten isopeptides. With the use of discovery-driven as well as targeted nanoscale liquid chromatography tandem mass spectrometry, we detected 29 TG2-gluten isopeptides in total, involving seven selected TG2 lysine residues (K205, K265, K429, K468, K590, K600, K677). Several gluten peptides carried known B-cell epitopes and/or T-cell epitopes, either intact 9-mer core regions or partial sequences, as well as sequences bearing striking similarities to already known epitopes. These novel insights into the molecular structures of TG2-gluten peptide complexes may help clarify their physiological relevance in the initiation of CD autoimmunity and the role of anti-TG2 autoantibodies.