Immunology ELISA Assay Kit

ENA screen ELISA Kit

$415.00

The ENA Screen (Extractable Nuclear Antigen) ELISA assay kit is used for the semi-quantitative determination of autoantibodies to nuclear and cytoplasmic antigens in human serum and plasma. The Eagle Biosciences ENA Screen (Extractable Nuclear Antigen) ELISA assay kit is for research use only and not to be used in diagnostic procedures.
This product was previously known as ENA31-K01.

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ENA screen ELISA Kit

ENA screen ELISA Kit Developed and Manufactured by Medipan

Size: 1×96 wells
Sensitivity: Cut-off Controls
Incubation Time: 2 hours
Sample Type: Serum, Plasma
Alternative Names: Extractable Nuclear Antigen screen ELISA, Human ENA screen ELISA, Human Extractable Nuclear Antigen screen ELISA
Sample Size: 100 µL
For Research Use Only

Reference Values
Positive: ≥ 1.0
Negative: < 1.0 It is recommended that each laboratory establishes its own normal and pathological reference ranges, as usually done for other diagnostic parameters, too. Therefore, the above mentioned reference values provide a guide only to values which might be expected. Sensitivity
The analytical sensitivity of each readivity of the ENA screen (Extractable Nuclear Antigen) ELISA is around 0.2.

Assay Principle

ENA screen (Extractable Nuclear Antigen) ELISA assay kit is an enzyme immunoassay for the semi-quantitative determination of IgG antibodies to nuclear and cytoplasmic antigens.  Antibodies of the calibrator and diluted samples react with nuclear and cytoplasmic antigens immobilized on the solid phase of microtiter plates. Recombinant SS-B, Sm, RNP (68 kDa, A, C), Scl-70, Jo-1 as well as highly purified Sm and SS-A guarantee the specific binding of autoimmune 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’-tetramethyl­benzidine (TMB) added into a blue product. The enzyme reaction is stopped by dispensing an acidic solution 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 cut-off is established by multiplying the OD of the calibrator with the corresponding factor.

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Additional Information

Assay Background

ENA screen (Extractable Nuclear Antigen) ELISA assay kit is used for the semi-quantitative determination of autoantibodies to nuclear and cytoplasmic antigens in human serum and plasma.  Systemic autoimmune diseases such as systemic lupus erythematosus, scleroderma, rheumatoid arthritis, Sjögren’s syndrome, dermatomyositis, mixed connective tissue disease are characterized by the appearance of a variety of autoantibodies directed against components of the cell nucleus.

ENA screen (Extractable Nuclear Antigen) ELISA assay kit allows the simultaneous detection of autoantibodies to the extractable nuclear antigens SS-A(Ro), SS-B(La), Sm, RNP and Scl-70 as well as the cytoplasmic antigen Jo-1 in one sample.  ENA screen (Extractable Nuclear Antigen) ELISA assay kit offers a rapid and handsome opportunity for the determination of the whole autoantibody pattern in systemic autoimmune diseases on one test plate. The use of specified recombinant antigens in combination with selected highly purified ones guarantees a maximum of specificity for these parameters.

Assay Procedure

  1. Bring all reagents to room temperature (18-25°C) before use. Mix gently without causing foam.
  2. Dispense 100 µl calibrator (Ca), 100 µl negative control (N), 100 µl diluted 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.

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