Anti-Gliadin sIgA / IgA ELISA Assay Kit

$680.00

SKU: GLI35-K01 Categories: , ,
Package insert view pdf

 

 

Intended Use
The Eagle Biosciences Anti-Gliadin sIgA / IgA ELISA Assay kit is intended for the quantitative determination of Anti-Gliadin sIgA / IgA in stool. The Anti-Gliadin sIgA / IgA ELISA kit is for research use only and not to be used in diagnostic procedures. 
 
Assay Principle
The Anti-Gliadin sIgA / IgA ELISA Assay kit 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 read at 450 nm in a microtiter plate reader. The anti-gliadin concentration can be calculated from the standard curve.

1.    Washing step

  • Take out the needed strips of the microtiter plate and wash 1x with 250 µl diluted WASHBUF.
  • Remove residual buffer by tapping the plate on absorbent paper after the washing step.

2.    Incubation samples

  • Pipette 100µl STD, CTRL and samples in duplicate in the microtiter plate. For standard 0 use the diluted WASHBUF
  • The strips are covered and incubated for 60 min at room temperature (18-26 °C).

3.    Washing step

  • Discard the content of the microwells and wash 5x with 250 µl diluted WASHBUF. Remove residual buffer by tapping the plate on absorbent paper after the last washing step.

4.    Incubation conjugate

  • Pipette 100 µl CONJ in each microwell.
  • The strips are covered and incubated for 60 min at room temperature (18-26 °C).

5.    Washing step

  • Discard the content of the microwells and wash 5x with 250 µl diluted WASHBUF.
  • Remove residual buffer by tapping the plate on absorbent paper after the last washing step.

6.    Incubation substrate

  • Pipette 100 µl SUB in each microwell.
  • Incubate for 10-15 min at room temperature (18-26 °C) in the dark.

7.    Stopping reaction

  • Pipette 50 µl STOP in each microwell. Mix well.

8.    Reading

  • Read the absorbance at 450 nm. If the microtiter plate reader allows to use a reference wavelength use 620 or 690 nm as reference wavelength.
  • Reading should be done within 5 min after stopping reaction.
  • If the highest standard exceeds the range of the reader the measurement should be done at 405 nm against 620 nm (690 nm).

 

Typical Standard Curve

anti-gliadin-iga.png

Assay Background
Celiac disease is a chronic illness of the small intestinal mucous membrane. It is caused by an intolerance against gluten, which is found in many cereals.  The intake of gluten-containing food leads to inflammation of the small intestinal mucous membrane. The resorption of nutrients is therefore reduced. The symptoms of the disease are reduction of weight, diarrhoea, vomiting, norexia and tiredness. The growth in children is reduced. The only therapeutic treatment is a gluten-free diet. 

An untreated celiac disease has been known to increase the risk of non-Hodgkin-lymphoma and colon cancer. In five to ten percent of the patients, celiac disease is associated with diabetes mellitus type 1. Women are more often affected than men. The outcome of the disease is pronounced during infancy and in an age between 30 and 40 years.

The Eagle Biosciences Anti-Gliadin sIgA / IgA ELISA Assay kit allows for an easy, rapid and precise quantitative determination of the secretory IgA and IgA gliadin antibodies in stool. The kit includes all reagents ready to use for preparation of the samples.