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Anti-Gliadin sIgA / IgA ELISA for Human Stool (ImmuChrom)

IC6300
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Description

The ImmuChrom Anti-Gliadin sIgA / IgA ELISA Kit is intended for the quantitative determination of anti-gliadin sIgA / IgA in stool. This immunoassay enables easy, rapid and precise quantitative determination of the antibodies in biological samples. For research use only.

Summary

Celiac disease is a chronic illness of the small intestinal mucous membrane. The cause is an intolerance against gluten, which is found in many cereals. The intake of gluten-containing food leads to an inflammation of the small intestinal mucous membrane, which results in a reduced absorption of nutrients. The symptoms of the disease are reduction of weight, diarrhea, vomitus, anorexia and tiredness. The growth of children is impaired. The characteristic of the symptoms might be different. The only therapeutic treatment is a gluten-free diet. A non-treated celiac disease increases the risk of non-Hodgkin-lymphoma and colon cancer. Celiac disease is associated with type 1 diabetes mellitus in five to ten percent of the patients. Women are more often affected than men. The outcome of the disease is pronounced during infancy and in ages between 30 and 40 years old.

Type

Sandwich ELISA, HRP-labelled antibody

Other names

secretory anti-gliadin immunoglobulin A, gliadin antibody, fecal anti-gliadin sIgA

Principle of Method

The anti-gliadin-ELISA test determines human anti-gliadin sIgA / IgA antibodies according to the “sandwich”-principle. Anti-gliadin antibodies in sample, standard and controls bind to gliadin, which is coated to the microtiter plate. After a washing step a peroxidase labeled detection antibody is added. A second washing step is followed by the addition of the substrate which is converted to a colored product by the peroxidase. The reaction is terminated by the addition of an acidic stop solution. The optical densities are measured at 450 nm (against the reference wavelength 620 nm) in a microtiter plate reader. The anti-gliadin concentration can be calculated from the standard curve.

Sample Types

Stool

Sample Volume

Extraction in Stool extraction vials
In a stool sample extraction vial mix 15 mg stool with 0.75 ml SAMPLEBUF, then vortex it until the mixture is homogenous. Transfer the resulting slurry to a plastic vial and centrifuge it for 10 min at 3000 xg. The supernatant is directly pipetted into the microtiter plate wells (100 µl in each well) with no further dilution.

Standard Curve

Example of a standard curve with the Anti-Gliadin sIgA / IgA ELISA kit for Human Stool (ImmuChrom)

Calibration Range:  25 - 200 mU/g

Limit of Detection

2.1 mU/g

For the determination the zero-standard was measured 20 times. The 2-fold standard deviation was added to the mean value of the optical density. The respective concentration was read from the standard curve.

Intra-assay (Within-Run) C.V.

  • 3.7% (21.5 mU/g) [n = 10]
  • 4.6% (47.0 mU/g) [n = 10]
  • 3.1% (79.7 mU/g) [n = 10]

Inter-assay (Run-to-Run) C.V.

  • 9.2% (22.5 mU/g) [n = 10]
  • 9.3% (48.0 mU/g) [n = 10]
  • 8.8% (83.5 mU/g) [n = 10]

Intended Use

This assay is intended for research use only and is not for use in diagnostic procedures.

Resources

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 Ilex Life Sciences LLC is an official distributor of Immuchrom GmbH products.