DESCRIPTION

C-reactive protein (CRP) is an evolutionary highly conserved member of the pentraxin superfamily of proteins. CRP consists of five identical subunits with molecular weight 20-30 kDa arranged as a cyclic pentamer. CRP exists in at least two conformationally distinct forms, native pentameric CRP (pCRP) and modified/monomeric CRP (mCRP). CRP plays important roles in inflammatory processes and host responses to infection including promoting agglutination, bacterial capsular swelling, phagocytosis and complement fixation through its Ca2+-dependent binding to phosphorylcholine (PCh). CRP is mainly expressed by hepatocytes and secreted into plasma. The concentration of CRP will increase significantly in plasma during acute phase response to tissue injury, infection or other inflammatory stimuli.

PRINCIPLE OF THE ASSAY

Sandwich ELISA structure (Detection antibody labeled with HRP) A capture antibody is pre-coated onto the bottom of wells which binds to analyte of interest. A detection antibody labeled with HRP also binds to the analyte. TMB acts as the HRP substrate and the solution color will change from colorless to blue. A stop solution containing sulfuric acid turns solution yellow. The color intensity is proportional to the quantity of bound protein which is measurable at 450 nm with the correction wavelength set at 630 nm.

PRECISION

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested 20 times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in 24 separate assays to assess inter-assay precision.

Intra-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	20	279.4	18.9	6.8
2	20	63.3	3.5	5.5
3	20	14.0	1.0	7.3

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	24	269.8	18.8	7.0
2	24	61.3	2.9	4.7
3	24	16.0	1.5	9.3

RECOVERY

The recovery of mouse C-Reactive Protein/CRP spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type		Average% of Expected	Range (%)
Mouse serum	1:320,000	111	107-114
	1:640,000	104	100-112
Cell culture supernatant	1:8	85	75-98
Tissue lysate	1:16	93	78-108

LINEARITY

To assess the linearity of the assay, samples were diluted with the appropriate Sample Diluent to produce samples with values within the dynamic range of the assay. (The mouse serum was initially diluted 1:40,000. The cell culture supernatant was initially diluted 1:2. )

		Mouse serum	Cell culture supernatant	Tissue lysate
1:2	Average% of Expected	100	100	100
	Range (%)	-	-	-
1:4	Average% of Expected	101	108	105
	Range (%)	95-106	105-112	91-119
1:8	Average% of Expected	92	112	79
	Range (%)	88-97	110-114	78-81
1:16	Average% of Expected	86	117	97
	Range (%)	82-89	115-119	93-101

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Mouse C-Reactive Protein/CRP Sandwich ELISA Kit produced at Sunlong Medical™.

Ordering & shipping

• Minimum order: 1 kit
• Lead time: varies with order quantity and specifications. Please consult your local distributor or our sales team for confirmation
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• Price: USD, per kit (48T / 96T options available)