Human C-Reactive Protein/CRP QuicKit ELISA

Catalog #: QK1707
1 Citation Datasheet / COA / SDS

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QK1707

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QC265: Human C-Reactive Protein/CRP QuicKit Control Group 265

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Citations (1)

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Summary Sample Values Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Human C-Reactive Protein/CRP QuicKit ELISA Summary

Assay Length

80 minutes

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL)

Sensitivity

0.079 ng/mL

Assay Range

0.8 - 50 ng/mL (Cell Culture Supernates, Serum, Plasma, EDTA Plasma, Heparin Plasma)

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.

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.

Product Summary

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.

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, Serum, EDTA Plasma, Heparin Plasma

	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

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

N/A

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Human CRP Standard Curve ELISA

ELISA

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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.

Product Datasheets

Product Datasheet

COA

SDS

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: C-Reactive Protein/CRP

C-Reactive Protein (CRP), also known as Pentraxin 1, is a non-glycosylated protein in the Pentraxin family that also includes Pentraxin 2/SAP and Pentraxin 3/TSG-14. CRP functions as a sensor and activator of the innate immune response (1). In humans, it is a major acute-phase protein; its circulating concentration is dramatically elevated at the onset of inflammation (2). In mice, however, serum CRP levels increase only slightly during inflammation, and the analogous acute phase role is filled by Pentraxin 2 (3). CRP assembles non-covalently into a 110-120 kDa cyclical pentamer (4). Mature human CRP shares 71% and 64% amino acid (aa) sequence identity with mouse and rat CRP, respectively (5).

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).

Entrez Gene IDs:

1401 (Human); 12944 (Mouse); 25419 (Rat)

Alternate Names:

C-Reactive Protein; C-reactive protein, pentraxin-related; CRP; MGC88244; pentraxin 1; PTX1MGC149895

Related Research Areas

Assay Procedure

These assays utilize an anti-tag coated microplate. Standards, samples, and controls are added to plate, followed by subsequent addition of an antibody cocktail. Following a 1 hour incubation, plates are washed and substrate is added and incubated for 20 minutes. Stop solution is then added and plates are read using a standard microplate reader.

QuicKit ELISA assay procedure

Citation for Human C-Reactive Protein/CRP QuicKit ELISA

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

Alveolar macrophages from EVALI patients and e-cigarette users: a story of shifting phenotype
Authors: Warren, KJ;Beck, EM;Callahan, SJ;Helms, MN;Middleton, E;Maddock, S;Carr, JR;Harris, D;Blagev, DP;Lanspa, MJ;Brown, SM;Paine, R;
Respiratory research
Species: Human
Sample Types: BALF