Anti-ASGPR (Asialoglycoprotein Receptor) ELISA Assay Kit

$340.00

The Eagle Biosciences Human Anti-ASGPR (Asialoglycoprotein Receptor) ELISA Assay Kit is used for the semi-quantitative determination of IgG antibodies to asialoglycoprotein receptor (ASGPR) in human serum or plasma. The Anti ASGPR (Asialoglycoprotein Receptor) ELISA Assay Kit is for research use only and not to be used in diagnostic procedures.

SKU: AAS31-K01 Categories: , ,

Anti-ASGPR (Asialoglycoprotein Receptor) ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: Cut-off Controls
Incubation Time: 2 hours
Sample Type: Serum, Plasma
Sample Size: 100 µL

Additional Information

Assay Background

Anti-ASGPR (Asialoglycoprotein Receptor) ELISA Assay Kit is used for the semi-quantitative determination of IgG antibodies to asialoglycoprotein receptor (ASGPR) in human serum or plasma.  ASGPR is a liver specific membrane receptor playing a pivotal role in the endocytosis of glycoproteins from the blood. An induction of humoral and cellular immune mechanisms to the ASGPR has been observed in the course of inflammatory liver disorders especially autoimmune hepatitis. The level of ASGPR autoantibodies correlates with the severity of the disease and declines under therapy.

The group of primary autoimmune liver disease (PAL) comprises autoimmune hepatitis (AIH), primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC).  Autoimmune hepatitis is a chronic inflammation of the liver with a yet unknown etiology. It comprises mild clinical forms as well as severe progressive hepatitis with lethal outcome. Females are more frequently affected. Clinical signs of the disease can occur as early as in their twenties.

Assay Principle

Anti-ASGPR (Asialoglycoprotein Receptor) ELISA Assay is an enzyme immunoassay for the semi-quantitative determination of IgG antibodies to ASGPR.   The antibodies of the calibrators and diluted samples react with ASGPR immobilized on the solid phase of microtiter plates. ASGPR highly purified from rabbit liver and coated on the microtiter plate guarantees the specific binding of ASGPR IgG antibodies of the specimen under investigation. Following an incubation period of 60 min at 37 °C, unbound serum components are removed by a washing step.

The bound IgG antibodies react specifically with anti-human-IgG conjugated to horseradish peroxidase (HRP) within an incubation period of 30 min at 37 °C. Excessive conjugate is separated from the solid-phase immune complexes by the following washing step.  Horseradish peroxidase converts the colorless substrate solution of 3,3’,5,5’-tetramethylbenzidine (TMB) added into a blue product. The enzyme reaction is stopped by dispensing an acidic solution (H2SO4) into the wells after 10 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.

Assay Procedure

  1. Bring all reagents to room temperature before use. Mix gently without causing foam.
  2. Dispense 100 µl controls (P, Co, N), 100 µl diluted patient samples, into the respective wells.
  3. Seal plate, incubate 60 min at 37 °C.
  4. Decant, then wash each well five times using 300 µl
    wash buffer (B).
  5. Add 100 µl of conjugate (D) solution to each well.
  6. Seal plate, incubate 30 min at 37 °C.
  7. Decant, then wash each well five times using 300 µl
    wash buffer (B).
  8. Add 100 µl of substrate (E) to each well.
  9. Incubate 10 min protected from light at room temperature.
  10. Add 100 µl of stop solution (F) to each well and mix gently.
  11. Read the optical density at 450 nm versus 620 or 690 nm within 30 min after adding the stop solution.

Manual

Product Manual