High Sensitive C-Reactive Protein (CRP) ELISA Kit

$695.00

The Eagle Biosciences High Sensitive C-Reactive Protein (CRP) ELISA Kit is intended for the quantification of C-Reactive Protein (CRP) in serum and plasma. The Eagle Biosciences CRP ELISA Assay Kit is for research use only and should not be used for diagnostic procedures.

SKU: ARG80870 Categories: , ,

High Sensitive C-Reactive Protein (CRP) ELISA Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 1 µg/mL
Dynamic Range: 5-100 µg/mL
Incubation Time: ~1 hour
Sample Type: Serum, Plasma
Sample Size: 10 µL

Additional Information

Assay Background

C-Reactive Protein (CRP) is an acute-phase protein, produced exclusively in the liver. Interleukin-6 is the mediator for the synthesis by the hepatocytes of CRP, a pentamer of approximately 120.000 Daltons. CRP is present in the serum of normal persons at concentrations ranging up to 5mg/l. The protein is produced by the fetus and the neonate and it does not pas the placental barrier, as such it can be used for the early detection of neonatal sepsis. Because febrile phenoena, leukocyte count and erytrhocyte sedimentation rate (ESR) are often misleading, investigators and clinicans now prefer a quantitative CRP determination as a marker for acute inflammation and tissue necrosis. Within 6 hours of an acute inflammatory challenge the CRP level starts to rise. Serum concentration of CRP increases significantly in cases of both infectious and non-infectious inflammation, of tissue damage and necrosis and in the presence of malignant tumours. CRP is present in the active stages of inflammatory disorders like rheumatoid arthritis, ankylosing spondylitis, Reiter’s syndrome, psoriatric arthropathy, systemic lupus erythematosus, polyarteritis, ulcerative colitis and Crohn’s disease. Injuries causing tissue breakdown and necrosis are associated with increases in serum CRP which are seen in thermal burns, major surgery and myocardial infarction. Widespread malignant disease with carcinoma of the lung, stomach, colon, breast, prostate and pancreas, Hodgkin’s disease, non-Hodgkin’s lymphoma and lymphosarcoma will give rise to high levels of CRP resulting from tissue damage by invading tumour cells. CRP, therefore may be used to monitor malignancy. The CRP-level increases dramatically following microbial infections, and this may be particularly helpful for the diagnosis and monitoring of bacterial septicemia in neonates and other immunocompromised patients at risk. In children, CRP is useful for differential diagnosis of bacterial and viral meningitis. Because the biological half-life of this protein is only 24 hours, CRP accurately parallels the activity of the inflammation process and the CRP concentration decreases much faster than ESR1,2 or any other acute phase parameter, which is particularly useful in monitoring appropriate treatment of bacterial diseases with antibiotics. C-Reactive Protein measurements during the early and late post transplant period of bone marrow and organ transplantations is particularly useful in the management of interfering infections in these immunosuppressed patients.

Assay Principle

The Eagle Biosciences High Sensitive C-Reactive Protein (CRP) ELISA Kit employs the quantitative sandwich enzyme immunoassay technique. An antibody specific for CRP has been pre-coated onto a microtiter plate. Standards or samples are pipetted into the wells and any CRP present in samples is bound by the immobilized antibody. After washing away any unbound substances, an HRP conjugated antibody specific for CRP is added to each well and incubate. A substrate solution (TMB) is then added to the wells and color develops in proportion to the amount of CRP bound in the initial step. The color development is stopped by the addition of acid and the intensity of the color is measured at a wavelength of 450 nm ±2 nm. The concentration of CRP in the sample is then determined by comparing the O.D of samples to the standard curve.

Assay Procedure

1. Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal it.
2. Add 100 μl of standards and samples in duplicate into wells.
3. Cover wells and incubate for 30 mins at RT.
4. Aspirate each well and wash, repeating the process 4 times for a total washes. Wash by filling each well with 1× Wash Buffer (350 μl) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating, decanting or blotting against clean paper towels.
5. Add 100 μl of HRP-conjugated antibody into each well.
6. Cover wells and incubate for 30 mins at RT.
7. Wash as according to step 4.
8. Add 100 μl of TMB Reagent to each well. Incubate for 10 minutes at room temperature in dark.
9. Add 50 μl of Stop Solution to each well. The color of the solution should change from blue to yellow.
10. Read the OD with a microplate reader at 450 nm immediately.

Manual

Product Manual