Description

Nori Caine CRP ELISA Kit Summary

Alternative names for CRP: C-reactive protein

Alternative name for canine: dog

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background:

C-reactive protein (CRP) is a protein found in the blood, the levels of which rise in response to inflammation (i.e. CRP is an acute-phase protein). CRP was first identified as a substance in the serum of patients with acute inflammation that reacted with the C-polysaccharide of Pneumococcus. Its physiological role is to bind to phosphocholine expressed on the surface of dead or dying cells (and some types of bacteria) in order to activate the complement system via the C1Q complex.^[1] CRP is synthesized by the liver^[2] in response to factors released by macrophages and fat cells (adipocytes).^[3] It is a member of the pentraxin family of proteins.^[2] C-reactive protein was the first pattern recognition receptor (PRR) to be identified.^[4] CRP rises up to 50,000-fold in acute inflammation, such as infection. It rises above normal limits within 6 hours, and peaks at 48 hours. Its half-life is constant, and therefore its level is mainly determined by the rate of production (and hence the severity of the precipitating cause).

CRP is used mainly as a marker of inflammation and infection. Measuring CRP level is a screen for infectious and inflammatory diseases. Rapid, marked increases in CRP occur with inflammation, infection, trauma and tissue necrosis, malignancies, and autoimmune disorders. Because there are a large number of disparate conditions that can increase CRP production, an elevated CRP level does not diagnose a specific disease. An elevated CRP level can provide support for the presence of an inflammatory disease, such as rheumatoid arthritis, polymyalgia rheumatica or giant-cell arteritis. However, CRP level is an independent risk factor for atherosclerotic disease. Patients with high CRP concentrations are more likely to develop stroke, myocardial infarction, and severe peripheral vascular disease.^[5]

Reference

1. Thompson D, Pepys MB, Wood SP (1999). Structure 7 (2): 169–77
2. Pepys MB, Hirschfield GM (June 2003). J. Clin. Invest. 111 (12): 1805–12.
3. Lau DC, Dhillon B, Yan H, et al. (May 2005). Am. J. Physiol. Heart Circ. Physiol. 288 (5): H2031–41.
4. Mantovani A, Garlanda C, Doni A, Bottazzi B (January 2008). J. Clin. Immunol. 28 (1): 1–13.
5. Clearfield MB (2005). The Journal of the American Osteopathic Association 105 (9): 409–16.

Product Citation