QuicKey Human CRP(C-Reactive Protein) ELISA Kit

SKU: E-TSEL-H0007

$ 331⁹⁵

QuicKey Human CRP(C-Reactive Protein) ELISA Kit

Detection Range	0.39-25 ng/mL
Sensitivity	0.23 ng/mL

Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T
Assay time	2.5h
Detection range	0.39-25 ng/mL
Sensitivity	0.23 ng/mL
Sample type &Sample volume	serum and plasma; 50μL
Specificity	This kit recognizes Human CRP in samples. No significant or interference between Human CRP and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of Human CRP concentrations in serum and plasma.Please consult technical support for the applicability if other biological fluids need to be tested.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human CRP. Samples (or Standards) and biotinylated detection antibody specific for Human CRP are added to the micro ELISA plate wells. Human CRP would combine with the specific antibody. Then Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human CRP, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 ± 2 nm. The OD value is proportional to the concentration of Human CRP. You can calculate the concentration of Human CRP in the samples by comparing the OD of the samples to the standard curve.

Kit Components and Storage

An unopened kit can be stored at 2-8℃ for six months. After test, the unused wells and reagents should be stored according to the table below.

Item	Specifications	Storage conditions after test
Micro ELISA Plate (Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	2-8℃, 1 month
Reference Standard	96T: 2 vials 48T: 1 vial	Discard unused reconstituted standard and dilutions
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8℃
Biotinylated Detection Ab Working Solution	1 vial, 6 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	2-8℃ (Protect from light)
Substrate Reagent	1 vial, 10 mL	2-8℃ (Protect from light)
Stop Solution	1 vial, 10 mL	2-8℃
Plate Sealer	5 pieces
Product Description	1 copy
Certificate of Analysis	1 copy

Technical Data:

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human CRP were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human CRP were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (ng/mL)	1.19	3.11	11.37	1.18	2.73	11.41
Standard deviation	0.07	0.15	0.55	0.06	0.13	0.51
CV (%)	5.88	4.82	4.84	5.08	4.76	4.47

Recovery

The recovery of Human CRP spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type	Range (%)	Average Recovery (%)
Serum(n=8)	95-103	98
EDTA plasma(n=8)	95-108	103

Target Information

Background

C-reactive protein (CRP) is a member of the phylogenetically ancient and highly conserved ‘pentraxin’ family of proteins, which also includes serum amyloid P component (SAP), a constituent of all amyloid deposits [1]. In man and other animal species, CRP is a major acute-phase plasma protein displaying rapid and pronounced rise of its serum concentration in response to infection or tissue injury.CRP is used mainly as a marker of inflammation. Apart from liver failure, there are few known factors that interfere with CRP production. Interferon alpha inhibits CRP production from liver cells which may explain the relatively low levels of CRP found during viral infections compared to bacterial infections [2]. Measuring and charting CRP values can prove useful in determining disease progress or the effectiveness of treatments. ELISA, immunoturbidimetry, nephelometry, rapid immunodiffusion, and visual agglutination are all methods used to measure CRP.
1. Gewurz, Henry. Biology of C-Reactive Protein and the Acute Phase Response [J]. Hospital Practice, 1982, 17(6):67-81.
2. Enocsson H , Christopher Sjwall, Skogh T , et al. Interferon-alpha mediates suppression of C-reactive protein: explanation for muted C-reactive protein response in lupus flares? [J]. Arthritis & Rheumatism, 2010, 60(12):3755-3760.

Database Links	SwissProt: P02741 Entrez Gene: 1401
Research Area	Cancer, Cardiovascular, Epigenetics and Nuclear Signaling, Metabolism, Immunology

Assay Procedures

	1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 90 min at 37°C
	2. Aspirate and wash the plate for 3 times
	3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	5. Add 50μL Stop Solution
	6. Read the plate at 450nm immediately. Calculation of the results