Human C-Reactive Protein/CRP QuicKit ELISA
R&D Systems, part of Bio-Techne | Catalog # QK1707
Key Product Details
Assay Length
80 minutes
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Serum (10 µL), EDTA Plasma (10 µL), Heparin Plasma (10 µL)
Sensitivity
0.079 ng/mL
Assay Range
0.781-50 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Product Summary for Human C-Reactive Protein/CRP QuicKit ELISA
The Quantikine® QuicKit™ Human C-Reactive Protein/CRP Immunoassay is a one step, 80-minute solid phase ELISA designed to measure human CRP levels in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant human CRP and antibodies raised against the recombinant protein. Results obtained using natural human CRP showed linear curves that were parallel to the standard curves obtained using the QuicKit™ standards. These results indicate that this kit can be used to determine relative mass values for natural human CRP.
Product Specifications
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Reactivity
Human
Specificity
Natural and recombinant human CRP.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Sample Values
Serum/Plasma - Samples from apparently healthy volunteers were evaluated for the presence
of human CRP in this assay. No medical histories were available for the donors used in this
study.
Cell Culture Supernates - Human peripheral blood mononuclear cells (PBMCs) (1 x 106 cells/mL) were cultured in RPMI supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. Cells were left unstimulated or stimulated with 10 μg/mL PHA for 5 days. Aliquots of the culture supernates were removed and assayed for levels of human CRP. No detectable levels were observed.
| Sample Type | Mean (ng/mL) | Range (ng/mL) | Standard Deviation (ng/mL) |
| Serum (n=10) | 2546 | 176-9946 | 2835 |
| ESTA plasma (n=20) | 2443 | 178-9519 | 2719 |
| Heparin plasma (n=20) | 2401 | 171-9550 | 2627 |
Cell Culture Supernates - Human peripheral blood mononuclear cells (PBMCs) (1 x 106 cells/mL) were cultured in RPMI supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. Cells were left unstimulated or stimulated with 10 μg/mL PHA for 5 days. Aliquots of the culture supernates were removed and assayed for levels of human CRP. No detectable levels were observed.
Precision
Intra-Assay Precision (Precision within an assay) Two samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Two samples of known concentration were tested in ten separate assays to assess inter-assay precision. Assays were performed by at least three technicians.
Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||
|---|---|---|---|---|
| Sample | 1 | 2 | 1 | 2 |
| n | 20 | 20 | 10 | 10 |
| Mean (ng/mL) | 3.75 | 26.1 | 4.03 | 24.8 |
| Standard Deviation | 0.146 | 1.08 | 0.195 | 1.31 |
| CV% | 3.9 | 4.1 | 4.8 | 5.3 |
Recovery for Human C-Reactive Protein/CRP QuicKit ELISA
The recovery of human CRP spiked to three levels in samples throughout the range of the assay was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 96 | 92-102 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human CRP were diluted with calibrator diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Human C-Reactive Protein/CRP QuicKit ELISA
Human CRP Standard Curve ELISA
Human CRP QuicKit Spiked Recovery Competitor Comparison
CRP is spiked at three known concentrations throughout the range of the assay and run to measure response of the spiked sample matrix. Culture media recovery is 95% compared to 84% for the top competitor. In spike and recovery experiments, natural samples are spiked with the recombinant target analyte of interest to identify interference caused by sample matrices.Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: C-Reactive Protein/CRP
CRP binds and opsonizes apoptotic cells (6-8) as well as bacteria such as S. pneumoniae (9, 10). It subsequently enhances the phagocytosis of these opsonized cells (6, 8-10). CRP additionally binds several proteins in the complement cascade including C1q, C4BP, and Factor H (8, 11-13). It enhances activation of the classical complement pathway and the deposition of C3b (9). In later stages of the response, CRP inhibits complement-mediated cell lysis through its binding to C4BP and Factor H (8, 12). These interactions induce the upregulation of complement inhibitory proteins CD46, CD59, and CD55/DAF and inhibit assembly of the membrane attack complex (MAC) (8, 14).
CRP binds to Fc gamma RI, Fc gamma RIIA, and Fc gamma RIIB on macrophages and dendritic cells (15-17), and Fc receptors are required for the phagocytosis of CRP-opsonized target cells (6, 10, 18). CRP binding to Fc gamma RI induces Src activation which subsequently triggers the inhibitory Fc gamma RIIb and dampens the inflammatory response (15, 19). CRP additionally promotes dendritic cell maturation and humoral immunity (10). In cardiovascular disease, CRP binds to oxidized LDL, exacerbates tissue damage in coronary artery infarction, and inhibits the repair of injured vascular endothelium (7, 19, 20).
Additional C-Reactive Protein/CRP Products
Product Documents for Human C-Reactive Protein/CRP QuicKit ELISA
Product Specific Notices for Human C-Reactive Protein/CRP QuicKit ELISA
For research use only
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