QuicKey Human CRP(C-Reactive Protein) ELISA Kit

    • QuicKey ELISA-Elabscience
    • ELISA-Elabscience
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    • QuicKey ELISA-Elabscience
    • ELISA-Elabscience
    • QuicKey ELISA-Elabscience
    • ELISA-Elabscience

      Catalog number:E-TSEL-H0007

      Size:
      • 96T
      Qty:
      - +
      Price: $390

      Reactivity: Human

      Detection Range: 0.39~25 ng/mL

      Sensitivity: 0.23 ng/mL

      Lead Time: 7~10 daysWelcome to order from local distributors.

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      QuicKey Series

      Get more sensitive and precise results with saving at least 1h comparing to traditional ELISA Kits. The new developed technology in house will help to accelerate your science research in a more efficient way.

       

      Intended use

      This ELISA kit applies to the in vitro quantitative determination of Human CRP concentrations in serum, plasma. Please consult technical support for the applicability if other biological fluids need to be tested.

       

      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. ELISAimmunoturbidimetrynephelometry, 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 Sjöwall, 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.

       

      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.

      Assay type

      Sandwich-ELISA

      Format

      96T/48T

      Assay time

      2.5h

      Reactivity

      Human

      Detection method

      Colormetric

      Detection range

      0.39-25 ng/mL

      Sensitivity

      0.23 ng/mL

      Sample volume

      50μL

      Sample type

      Serum, Plasma



      Specificity

      This kit recognizes Human CRP in samples. No significant cross-reactivity or interference between CRP and analogues was observed.


      Typical data

      As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

      Concentration(ng/mL)

      25

      12.5

      6.25

      3.13

      1.56

      0.78

      0.39

      0

      OD

      2.434

      1.587

      1.027

      0.607

      0.353

      0.211

      0.134

      0.068

      Corrected OD

      2.366

      1.519

      0.959

      0.539

      0.285

      0.143

      0.066

      -

       


      Sample values

      Serum/Plasma–Samples from apparently healthy volunteers were evaluated for the presence of Human CRP in this assay.

      Sample Type

      Source

      Range

      Dilution Factor

      Serum(n=12)

      Healthy human

      0.21-4.53μg/mL

      50-4000

      EDTA plasma(n=12)

      Healthy human

      0.16-4.72μg/mL

      20-4000

       

      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=13)

      95-103

      98

      EDTA plasma (n=13)

      95-108

      103

       

      Linearity

      Samples were spiked with high concentrations of Human CRP and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

       

       

      Serum (n=4)

      EDTA  plasma (n=4)

      1:2

      Range (%)

      91-103

      94-103

      Average (%)

      96

      98

      1:4

      Range (%)

      95-105

      98-109

      Average (%)

      99

      102

      1:8

      Range (%)

      91-104

      89-96

      Average (%)

      99

      92

      1:16

      Range (%)

      95-105

      83-96

      Average (%)

      99

      91

       

      Kit components & 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

      Stop Solution

      1 vial, 10 mL

      2-8℃

      Plate Sealer

      5 pieces

       

      Product Description

      1 copy

      Certificate of Analysis

      1 copy

      Note: All reagent bottle caps must be tightened to prevent evaporation and microbial pollution.

      The volume of reagents in partial shipments is a little more than the volume marked on the label, please use accurate measuring equipment instead of directly pouring into the vial(s).


      Other supplies required

      Microplate reader with 450 nm wavelength filter

      High-precision transfer pipette, EP tubes and disposable pipette tips

      37℃ Incubator

      Deionized or distilled water

      Absorbent paper

      Loading slot for Wash Buffer


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