Anti-dsDNA ELISA Assay Kit

$350.00

The Anti-dsDNA ELISA Assay Kit is used for the quantitative determination of IgG antibodies to double-stranded desoxyribonucleic acid (dsDNA) in human serum for the diagnosis of systemic lupus erythematosus (SLE). The Eagle Biosciences Anti-dsDNA ELISA is for research use only and not for diagnostic procedures.

SKU: DSD31-K01 Categories: , ,

Anti-dsDNA ELISA Assay Kit

Anti-dsDNA ELISA Assay Kit Developed and Manufactured by Medipan

Size: 1×96 wells
Sensitivity: 1 IU/ml
Dynamic Range: 1 – 300 IU/ml
Incubation Time: 2.5 hours
Sample Type: Serum, Plasma
Alternative Names: Anti dsDNA ELISA, Human Anti-dsDNA ELISA
Sample Size: 10 µL

For Research Use Only


Specificity
Testing 102 non-selected sera from blood donors a specificity of 99% was determined in Anti-dsDNA.


Assay Principle for Anti-dsDNA ELISA

The Anti-dsDNA ELISA Assay is an enzyme immunoassay for the quantitative determination of IgG antibodies to dsDNA.  The antibodies of the calibrators, control and diluted patient samples react with dsDNA immobilized on the solid phase of microtiter plates. Highly purified dsDNA coated on the microtiter plate guarantees the specific binding of dsDNA IgG antibodies of the specimen under investigation. Following an incubation period of 60 min at room temperature (RT), unbound sample 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 into the wells after 15 minutes at room temperature turning the solution from blue to yew. 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 antibody concentration of the specimen is directly read off the standard curve.  Alternatively, results can be calculated by a semi-quantitative method too using calibrator 2 as cut-off calibrator.


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Additional Information

Assay Background


Systemic autoimmune diseases such as SLE are characterized by the appearance of a variety of autoantibodies directed against cell components of the nucleus or plasma. Although significance and pathological relevance of some autoanti¬bodies are not completely revealed yet, the detection of autoantibodies is widely established and plays an important role in the diagnostics of systemic autoimmune diseases.

SLE has an unknown etiology and is characterized by multiorgan pathology. SLE has a female predominance. The onset of the disease occurs usually during childbearing age.

Antibodies to dsDNA are the hallmark for SLE diagnostics and are included in the diagnostic criteria of the American College of Rheumatology for SLE (1,2).

Assay Procedure


  1. Bring all reagents to room temperature (18-25°C) before use. Mix gently without causing foam.
  2. Dispense  100 µl calibrators 0 (optionally) 1 – 4 or 100 µl calibrator 2 (semi-quantitative) 100 µl controls P, N (N optionally) 100 µl diluted patient samples into the respective wells.
  3. Cover plate, incubate 60 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) solution to each well.
  6. Cover plate, 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 15 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


DSD31-K01 Anti dsDNA ELISA Assay Kit

Manual

Product Manual