Immunology ELISA Assay Kit

Anti-SLA ELISA Assay Kit

$565.00

The Anti-SLA ELISA Assay Kit is used for the quantitative determination of IgG antibodies to soluble liver antigen (SLA) in human serum or plasma for the diagnosis of autoimmune hepatitis (AIH). The Eagle Biosciences Anti-SLA ELISA is for research use only and not for diagnostic procedures.
This product was previously known as SLA31-K01.

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Anti-SLA ELISA Assay Kit

Anti-SLA ELISA Assay Kit Developed and Manufactured by Medipan

Size: 1×96 wells
Sensitivity: 3 U/ml
Dynamic Range: 3 – 300 U/ml
Incubation Time: 2 hours
Sample Type: Serum, Plasma
Sample Size: 10 µL
Alternative Names: Soluble Liver Antigen ELISA, Human Anti-SLA ELISA
For Research Use Only

Assay Background

The group of primary autoimmune liver disease (PAL) comprises AIH, primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC). The clinical picture of PAL is in most cases not different from other chronic liver diseases. About 15% of all cases with chronic liver diseases show an autoimmune pathogenesis. Therefore, after exclusion of infectious aetiology especially by viruses, the determination of different autoantibodies is recommended. Patients suffering from AIH show a variety of autoantibodies. Due to the appearance of different antibody specificities classification of AIH into different subgroups is discussed. Type I is characterized by the occurrence of antinuclear antibodies (ANA) and antibodies to smooth muscles (ASMA). ASMA recognize antigenic structure formed by polymeric f-Actin. For type II a high prevalence of antibodies to liver and kidney microsomal antigens (LKM) has been described. LKM1 antibodies recognize epitopes of cytochrome P450 IID6, a 50 kDa cyto¬plasmic protein found in hepatocytes and proximal tubular kidney cells. LC1 antibodies are specific for type II hepatitis, too. The respective antigen is formiminotransferase / cyclodeaminase located in the cytosol of liver cells. Patients with type III autoimmune hepatitis exhibit antibodies to the soluble liver antigen (SLA) and to the liver pancreas antigen (LP). It has been shown that both antibody species recognise the same antigen and thus are identical. The yet unknown antigen is suggested to be an UGA suppressor tRNA-associated protein. However, type III AIH is not yet fully accepted as independent subgroup for autoimmune hepatitis by the “International Hepatitis Group”. SLA is found in up to 20% of AIH cases and very often the only autoantibody associated with AIH to be detected.

Products Related to Anti-SLA ELISA Assay

Anti-LKM1 ELISA Assay Kit
Anti-M2 ELISA Assay Kit
VDBP ELISA Assay Kit

Additional Information

Assay Principle

The Eagle Biosciences Anti-SLA ELISA Assay Kit is an enzyme immunoassay for the quantitative determination of IgG antibodies to SLA.  The antibodies of the calibrators, controls, and diluted patient samples react with SLA immobilized on the solid phase of microtiter plates. The use of human recombinant SLA guarantees the specific binding of SLA antibodies of the specimen under investigation. Following an incubation period of 30 min at room temperature (RT), unbound serum components are removed by a wash step. The bound IgG antibodies react specifically with anti-human-IgG conjugated to horseradish peroxidase (HRP) within the incubation period of 30 min at RT. 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. The enzyme reaction is stopped by dispensing an acidic solution (HCl) into the wells after 30 min at RT 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 antibody concentrations 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.

Assay Procedure

  1. Bring all reagents to room temperature (18-25°C) before use. Mix gently without causing foam.
  2. Dispense  100 µl calibrators (1 – 6), 100 µl positive (P) and negative (N) control, 100 µl diluted patient samples into the respective wells.
  3. Incubate 30 min at room temperature (18-25°C).
  4. Decant, then wash each well three times using 300 µl  wash solution (made of B).
  5. Add 100 µl of conjugate (D) to each well.
  6. Incubate 30 min at room temperature (18-25°C).
  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 30 min protected from light at room temperature (18-25°C).
  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-SLA ELISA Standard Curve

Manual

Product Manual

Publications

Literature

  • Czaja AL. Natural history, clinical features and treatment of autoimmune hepatitis. Semin Liver Dis (1987) 4: 1 – 12
  • Johnson PJ, McFarlane IA. Meeting Report: International Autoimmune Hepatitis Group. Hepatology (1993) 18: 998 – 1005

Citations