Celiac EmA (Anti-Endomysium Antibody) IgG ELISA Assay kit: For Research Use Only Size: 1x96 wells Sensitivity: 1.0 U/ml Dynamic Range: 1 - 300 U/ml Incubation Time: 2 hours Sample Type: Serum Sample Size: 100 µl Controls Included
Intended Use The Celiac EmA (Anti-Endomysium Antibody) IgG ELISA assay kit is used for the quantitative determination of IgG autoantibodies to sample endomysial autoantigens (EmA). The Eagle Biosciences Celiac EmA (Anti-Endomysium Antibody) IgG ELISA assay kit is for research use only and not to be used in diagnostic procedures.
Assay Principle Celiac EmA (Anti-Endomysium Antibody) IgG ELISA assay kit is an enzyme immunoassay for the quantitative determination of IgG autoantibodies to the supposed major autoantigens in Dermatitis herpetiformis and celiac disease in sample serum.
Autoantibodies of the diluted samples and standards react with the immobilized human autoantigens on the solid phase of the microtiter plate. Celiac EmA (Anti-Endomysium Antibody) IgG ELISA assay kit guarantees the specific binding of autoantibodies usually found by immunofluorescence on endomysial antigens. Following an incubation period of 60 min at room temperature (18…25°C), unbound serum components are removed by a wash step.
The bound autoantibodies react specifically with anti-human-IgG-antibodies conjugated to horseradish peroxidase (HRP) within the incubation period of 30 min at room temperature. Excessive conjugate is separated from the solid-phase immune complexes by the following wash step.
HRP converts the colorless substrate solution of 3,3’,5,5’-tetramethylbenzidine (TMB) added into a blue product. This enzyme reaction is stopped by dispensing an acidic solution (H2SO4) into the wells after 15 min at room temperature turning the solution from blue to yellow.
The optical density (OD) of the solution at 450 nm is directly proportional to the amount of specific antibodies bound. The standard curve is established by plotting the concentrations of the antibodies of the calibrators (x-axis) and their corresponding OD values (y-axis) measured. The concentration of antibodies of the specimen is directly read off the standard curve. Evaluating the test by a semi-quantitative method is also possible.
Typical Standard Curve
Assay Background: Several autoantigens have been discussed for celiac disease and Dermatitis herpetiformis recently. Dieterich et al. have described two non-collagenous proteins with an apparent molecular weight of 90 and 300 kDa being the main autoantigens detected by EmA (1). Celiac disease, or gluten-sensitivity, is found already in neonates and is characterized by small intestinal damages leading to a so-called “flat” mucosa. Due to this extensive lesions mal-absorption occurs frequently accompanied with a depletion of key nutrients.
Incidence rates for celiac disease range from 1 in 300 (Western Ireland) to 1 in 4700 in European countries. However, a high number of subclinical cases of celiac disease have been detected by in-vitro tests revealing a prevalence of 4 in 1000. Individuals suffering from prolonged celiac disease additionally face an elevated risk of developing T cell lymphoma. Dermatitis herpetiformis is an autoimmune chronic relapsing skin disease characterized by formation of sub-epidermal blisters. Deposits of IgG antibodies are found in the basal membrane associated with neutrophil and eosinophil infiltration. Dermatitis herpetiformis seems to demonstrate celiac like changes in the intestine resembling the inflammatory process in celiac samples.