Anti-tTG IgG ELISA Kit

$470.00

The Anti-tTG IgG ELISA Assay Kit is used for the quantitative or semi-quantitative determination of IgG autoantibodies to tissue transglutaminase (tTG) in human serum or plasma. The Eagle Biosciences Anti-hu tTG IgG ELISA is for research use only and not for diagnostic procedures.

SKU: TGG31-K01 Categories: , ,

Anti-tTG IgG ELISA Kit

Anti-tTG IgG ELISA Kit Developed and Manufactured in the USA

Size: 1×96 wells
Sensitivity: 1 U/ml
Dynamic Range: 1 – 300 U/ml
Incubation Time: 2.5 hours
Sample Type: Serum, Plasma
Sample Size: 10 µL
Alternative Names: Anti-Human tTG IgG ELISA, Human Anti-Tissue Transglutaminase IgG ELISA
For Research Use Only


Assay Principle


The Anti-tTG IgG ELISA Kit ELISA Kit is an enzyme immunoassay for the quantitative or semi-quantitative determination of IgG autoantibodies to tissue transglutaminase in human serum or plasma.

Autoantibodies of the diluted patient samples, positive control, and calibrators react with human tissue transglutaminase immobilized on the solid phase of a microtiter plate. Anti-huTransG guarantees the specific binding of anti-tTG IgG autoantibodies of the specimen under investigation by employing highly purified, activated recombinant human tTG for coating. 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’-tetramethyl¬benzidine (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.


Related Products

Anti-tTG IgA ELISA Kit
Celiac EmA IgA ELISA
Celiac EmA (Anti-Endomysium Antibody) IgG ELISA Assay Kit

Additional Information

Assay Background


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. Gliadin the alcohol soluble fraction of gluten represents the causative agent of celiac disease that provokes an inflammatory process in the small intestine. Gliadin is a substrate of tTG and cross-linked into high molecular complexes triggering probably both cellular and humoral immune responses.

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.

Diagnosis of celiac disease comprises small intestine biopsy demonstrating a “flat” mucosa prior to a gluten-free diet and the following reconstitution of the mucosa after onset of the diet. Determination of anti-gliadin IgG and IgA by ELISA as well as the detection of anti-endomysium IgA by immunofluorescence has been considered as the main serological parameters for celiac disease so far.

The identification of tissue transglutaminase as one of the main endomysial autoantigens and the availability of an easy to use and reliable ELISA kit employing recombinant human tissue transglutaminase pro¬mises the extension of diagnostic opportunities for celiac disease in future.

IgA deficient celiac patients may demonstrate IgG autoantibodies to tTG only. Therefore, tTG IgG autoantibodies are the only serological parameter for celiac disease detectable in such patients apart from Gliadin IgG antibodies.

Assay Procedure


  1. Bring all reagents to room temperature (18-25°C) before use. Mix gently, avoid foam.
  2. Dispense 100 µl calibrators (0 optional) 1 – 4 (quantitative) or 100 µl calibrator 1 (semi-quantitative) 100 µl control P (N optional) 100 µl diluted patient samples into the respective wells.
  3. Seal plate, incubate 60 min at room temperature.
  4. Decant, then wash each well three times using 300 µl wash solution (made of B).
  5. Add 100 µl of conjugate (D) solution to each well.
  6. Seal plate, incubate 30 min at room temperature.
  7. Decant, then wash each well three times using 300 µl wash solution (made of B).
  8. Add 100 µl of substrate (E) to each well.
  9. Incubate 15 min protected from light at room temperature.
  10. Add 100 µl of stop solution (F) to each well and mix gently.
  11. Read the OD at 450 nm versus 620 or 690 nm within 30 min after adding the stop solution

Typical Standard Curve


Anti-tTG IgG ELISA Kit Standard Curve

Manual

Product Manual


Publications

Literature


  • Schuppan D, Hahn EG: IgA anti-tissue transglutaminase: setting the stage for celiac disease screening. Eur J Gastroenterol Hepatol. 2001 13 635-7.
  • Dieterich W, Ehnis T, Bauer M, Donner P, Volta U, Riecken EO, Schuppan D: Identification of tissue transglutaminase as the autoantigen of celiac disease. Nat Med 1997 3 797-801.