Immunology ELISA Assay Kit

Anti-CCP ELISA Assay Kit

$565.00

The Anti-CCP ELISA Assay Kit is designed and validated for the quantitative determination of IgG antibodies against cyclic citrullinated peptides (CCP) in human serum. The Eagle Biosciences anti-CCP ELISA Assay Kit is for research use only and should not be used in diagnostic procedures. 

SKU: CCP31-K01 Categories: , ,

Anti-CCP ELISA Assay Kit

Anti-CCP ELISA Assay Kit Developed and Manufactured by Medipan

Size: 1×96 wells
Sensitivity: 1.2 U/mL
Dynamic Range: 1 – 2000 U/mL
Incubation Time: 2 hours
Sample Type: Serum, Plasma
Sample Size: 5 µL
Alternative Names: Human Anti-CCP ELISA, Human CCP IgG ELISA
For Research Use Only

Reference Values
Negative: < 30 U/ml Positive: ≥ 30 U/ml It is recommended that each laboratory establishes its own normal and pathological reference ranges for serum anti-CCP antibodies levels as usually done for other diagnostic parameters, too. Therefore, the above mentioned reference values provide only a guide.

Assay Background

Rheumatoid Arthritis (RA) is most often diagnosed by the measurement of rheumatoid factors (RF). The rheumatoid factor is an antibody directed against the Fc-region of IgG. It appears mainly as IgM antibody but also as IgA or IgG subgroup. Rheumatoid factors are present in sera of 70 – 80 % of patients suffering from rheumatoid arthritis. But it is not specific for RA since it will also be found in healthy persons. The incidence depends on the age: for young people it is 1 – 4 %, for older persons it may reach up to 25 %.
The advantage of CCP antibodies is a higher sensitivity and specificity for the diagnosis of rheumatoid arthritis in comparison to the rheumatoid factors alone. Anti-CCP is often found at a very early state of the disease and it has a high predictive value for development of the disease.

Related Products

RF IgG (Rheumatoid Factor) ELISA Assay Kit
RF IgM (Rheumatoid Factor) ELISA Assay Kit
RF (Rheumatoid Factor) IgA ELISA Assay Kit

Additional Information

Assay Principle

Medizym® anti-CCP Ref is an enzyme immunoassay for the quantitative determination of IgG autoantibodies to cyclic citrullinated peptides (CCP) in human serum.

In the first step CCP AAb from the diluted sample (as well as from the calibrators and control) bind to cyclic citrullinated peptides coated on the microtiter plate. After an incubation of 60 minutes at room temperature (RT) unbound components are removed by washing step.

In a next step bound antibodies reacts with the added anti-human-IgG horseradish peroxidase (HRP) complex. Excessive conjugate is removed after 30 minutes at RT by another washing step.

HRP converts the colorless substrate TMB added into a blue product. The enzyme reaction is stopped by adding an acid solution after 15 minutes at RT. The color changes from blue to yellow. The absorbance of the resulting product is measured at 450 / 620 nm within 30 minutes. The obtained OD is direct proportional to the amount of bound antibodies.

Assay Procedure

  1. Pipette into the corresponding wells according to assay scheme
  2. 100 µl calibrators (1 – 5)
  3. 100 µl diluted patient sample and control serum (C).
  4. Cover the plate and incubate for 60 min at RT (18 – 25 °C).
  5. Aspirate or “flick out” by striking the wells sharply onto absorbent paper to remove any residual droplets. Wash 3 times with 300 µl washing solution (diluted from B) with 5 seconds soaking time each.
  6. Add 100 µl of anti-human IgG – HRP (D) to each well.
  7. Cover the plate and incubate for 30 min at RT.
  8. Aspirate or “flick out” by striking the wells sharply onto absorbent paper to remove any residual droplets. Wash 3 times with 300 µl washing solution (diluted from B) with 5 seconds soaking time each.
  9. Add 100 µl substrate solution (E) to each well and shake shortly.
  10. Incubate for 15 min in the dark at RT.
  11. Add 100 µl stop solution (F) to each well.
  12. Avoid any time shift during pipetting the samples and reagents.
  13. Read the optical density (OD) at 450 nm versus 620 or 690 nm within 30 min after adding the stop solution.

Documents

Publications

Citations

Selvam SP, Chinnadayyala SR, Cho S. Electrochemical Nanobiosensor for Early Detection of Rheumatoid Arthritis Biomarker: Anti-Cyclic Citrullinate Peptide Antibodies Based on Polyaniline (PANI)/MoS2-Modified Screen-Printed Electrode With PANI-Au Nanomatrix-Based Signal Amplification. Sens Actuators B Chem. 2021;333:129570.

Al-Sayed, MT;Mohammed, AS;Mohammed, AH;Moshrif, AM;, (2017). Diagnostic and prognostic values of serum HSP70 and YKL-40 in patients with rheumatoid arthritis. International Journal of Clinical Rheumatology.