Human C-Reactive Protein (CRP) ELISA KIT

Sample

Serum

Species Reactivity

Human

Detection Method

sELISA

Intended Use

For quantitative detection of C-Reactive Protein (CRP) in serum. For research use only.

Contents of Kit

1. Anti-human CRP coated strip plate: 96 wells
2. CRP Std. A (0 ng/ml), or Sample Diluent: 16 ml
CRP Std B (100 ng/ml): 0. 50 ml
CRP Std C (400 ng/ml): 0. 50 ml
CRP Std D (1000 ng/ml): 0. 50 ml
CRP Std E (4000 ng/ml): 0. 50 ml
CRP Std F (10,000 ng/ml): 0. 50 ml
3. Human CRP Low & High Control in a buffer: 0.5 ml each, (Lot sp. Conc given on the vial)
4. Anti-hCRP-HRP Conjugate: 0.3ml,Dilute 1:80 with assay buffer
5. Assay Buffer: 40 ml
6. HRP substrate, Solution: 16 ml
7. Wash buffer (10×): 50 ml; dilute 1:10 with distilled water
8. Stop solution: 6 ml
9. Instruction Manual

Storage

The microtiter well plate and all other reagents, if unopened, are stable at 2-8°C until the expiration date printed on the label. The whole kit stability is at least 6 months from the date of shipping under appropriate storage conditions. After opening the kit components, the shelf life is approx. 2 months.

Performance Characteristics

HIGH DOSE HOOK EFFECT: CRP concentrations of up to 160, 000 ng/ml did not show any hook effect.

Precision

Intra-assay precision: Three serum samples (mean CRP concentrations 205.8, 769.2, 8437.8 ng/ml) were run in 10 replicates. The samples showed good intra-assay precision with %CV of 12, 5, and 6.3, respectively.
Inter-assay precision: Three serum samples (227, 1022.2, 8791.8 ng/ml) were run in duplicate in sixteen independent assays. The samples showed good inter-assay precision (9.9, 9.5, and 7.8% CV).

Detection Limit

Based on sixteen replicate determinations of the zero standard, the minimum CRP concentration detectable using this assay is 10 ng/ml. The detection limit is defined as the value deviating by 2 SD from the zero standard.

General Description

C-reactive protein (CRP) has been regarded as an acute phase reactant in serum. It consists of five single subunits, which noncovalently linked and assembled, as a cyclic pentamer with a mol. Wt. Range of 110-140 kDa. CRP has been found to be increased in serum of patients with a wide variety of diseases including infections by gram-positive and gram-negative bacteria, acute phase of rheumatoid arthritis, abdominal abscesses, inflammation of bile ducts, myocardial infarction, and malignant tumors. CRP may be found in patients with Guillain-Barre syndrome and multiple sclerosis, certain viral infections, tuberculosis, acute infectious hepatitis, many other necrotic and inflammatory diseases, burned patients, and after surgical trauma. Although the detection of elevated levels of CRP in the serum is not specific for any particular disease, it is useful indicator of inflammatory processes. CRP levels rise in serum within hours of the onset of inflammation, reach a peak during the acute stage and decrease with resolution of inflammation trauma. The detection of CRP is a more reliable and sensitive indicator of the inflammatory process than the erythrocyte sedimentation rate, which may also be influenced by physiological changes not associated with an inflammation process. Current quantification methods including latex agglutination, nephelometry, radial immunodiffusion have the general disadvantage accompany agglutination and precipitation techniques.

Standard Curve

NOTE: These data are for demonstration purpose only. A complete standard curve must be run in every assay to determine sample values. Each laboratory should determine their own normal reference values.