Anti-GP2 (Glycoprotein 2) IgA ELISA Assay Kit

$340.00

The Eagle Biosciences Anti-GP2 IgA ELISA Assay kit is used for the quantitative determination of IgA antibodies against glycoprotein 2 (GP2) in human serum for the diagnosis of Crohn’s disease. The Anti-GP2 IgA is for research use only and not for use in diagnostic procedures.

SKU: G2A31-K01 Categories: , ,

Anti-GP2 (Glycoprotein 2) IgA ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 1 U/ml
Dynamic Range: 1 – 300 U/ml
Incubation Time: 1.5 hours
Sample Type: Serum, Plasma
Sample Size: 10 µL

Additional Information

Assay Background

Non-specific inflammatory bowel disease including Crohn’s disease (Enteritis regionalis) and ulcerative colitis (UC) are characterised by unknown etiology as well as chronic-remitting inflammatory processes of the intestine. Whereas the inflammation of ulcerative colitis is restricted to the mucosa and submucosa of colon and rectum, Crohn’s disease (CD) shows a wide spread inflammation of gastro-intestinal tract with granuloma formation. The risk developing one of these diseases is strongly correlated to immunologic, genetic, infectious and environmental factors.

The differential diagnosis of inflammatory bowel diseases to chronic diarrhea, recurrent abdominal dolor, infectious colitis, anorexia as well as the differentiation of CD to UC is still a challenge.

Autoantibodies of exocrine pancreas (PAB) were identified as specific serological marker for CD. A prevalence of 39 % of these autoantibodies in patients with CD could be demonstrated by indirect immune fluorescence (Stöcker et al. 1987). Glycoprotein 2 (GP2), a membrane-bound pancreatic protein, could be identified/verified as the major target of PAB’s (Roggenbuck et al., 2009). In combination with the detection of autoantibodies to Saccharomyces cerevisiae (ASCA) with a prevalence of 70 % in patients with CD and atypical antineutrophile cytoplasmatic antigens (aANCA) which are mainly found in patients with UC, PAB’s against GP2 could be used as a highly specific serological marker for differential diagnosis of CD to UC.

Assay Principle

The Eagle Biosciences Anti-GP2 IgA is an enzyme immunoassay for the quantitative determination of IgA antibodies to glycoprotein 2.

The antibodies of the calibrators, positive control, and diluted patient samples react with the antigen immobilized on the solid phase of microtiter plates. After an incubation period of 60 min at room temperature (18…25°C), unbound serum components are removed by a wash step.

The bound IgA autoantibodies react specifically with anti-human-IgA-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.

Typical Standard Curve

Manual

Product Manual


Publications

Literature

  • Stöcker W, Otte M, Ulrich S, Normann D, Finkbeiner H, Stöcker K, Jantschek G, Scriba PC: Autoimmunity to pancreatic juice in Crohn’s disease. Results of an autoantibody screening in patients with chronic inflammatory bowel disease. Scand J Gastroenterol Suppl. 1987; 139: 41-52.
  • Roggenbuck D, Hausdorf G, Martinez-Gamboa L, Reinhold D, Büttner T, Jungblut PR, Porstmann T, Laass MW, Henker J, Büning C, Feist E, Conrad K: Identification of GP2, the major zymogen granule membrane glycoprotein, as the autoantigen of pancreatic antibodies in Crohn’s disease. Gut, 2009;58:1620-8.