QuicKey Human CLU(Clusterin) ELISA Kit

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

      Catalog number:E-TSEL-H0014

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

      Reactivity: Human

      Detection Range: 1.56~100 ng/mL

      Sensitivity: 0.94 ng/mL

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

      Add to cart Compare Bulk request

      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 CLU concentrations in serum, plasma, urine, saliva. Please consult technical support for the applicability if other biological fluids need to be tested.


      Background

      Clusterin is a ubiquitous protein that is arousing increasing interest owing to its widespread diffusion and multifunctional role. It is composed of two 40 kD subments (NA1, NA2) encoded by a single gene, which hold together by disulphide bonds. The mature dimer has a molecular mass of 80 kD and is micro-heterogeneous in electrical charge due to glycosylation. This protein has been differently named as apolipoprotein J (ApoJ), sulfated glycoprotein 2 (SGP-2), glycoprotein III (GP- III), testosterone repressed prostate message (TRPM-2), SP-40, 40cytolysis inhibitor (CL1) and others [1].It is a component of the high-density lipoprotein (HDL) complex with putative functions in the transport of lipids, apoptosis, protection of cells from stress and, most important, in the regulation of complement activity. It has been postulated that clusterin is involved in a variety of human diseases including Alzheimer disease, brain injury following ischemianephrosis and myocarditis [2].


      1. Wilson M R, Easterbrooksmith S B. Clusterin is a secreted mammalian chaperone. [J]. Trends in Biochemical Sciences, 2000, 25(3):95-98.

      2. Mclaughlin L, Zhu G, Mistry M, et al. Apolipoprotein J/clusterin limits the severity of murine autoimmune myocarditis. [J]. Journal of Clinical Investigation, 2000, 106(9):1105-1113.


      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 CLU. Samples (or Standards) and biotinylated detection antibody specific for Human CLU are added to the micro ELISA plate wells. Human CLU would combined 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 CLU, 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 CLU. You can calculate the concentration of Human CLU in the samples by comparing the OD of the samples to the standard curve.

      Assay type

      Sandwich-ELISA

      Format

      96T

      Assay time

      2.5h

      Reactivity

      Human

      Detection method

      Colormetric

      Detection range

      1.56-100 ng/mL

      Sensitivity

      0.94 ng/mL

      Sample volume

      100μL

      Sample type

      Serum, Plasma, Urine, Saliva


      Specificity

      This kit recognizes Human CLU in samples. No significant cross-reactivity or interference between CLU 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)

      100

      50

      25

      12.5

      6.25

      3.13

      1.56

      0

      OD

      2.402

      1.527

      0.912

      0.539

      0.319

      0.196

      0.128

      0.062

      Corrected OD

      2.340

      1.465

      0.850

      0.477

      0.257

      0.134

      0.066

      -






      Sample values

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

      Sample Type

      Source

      Range

      Dilution Factor

      Serum (n=14)

      Healthy human

      61-346μg/mL

      4000-32000

      EDTA plasma (n=11)

      Healthy human

      159-380μg/mL

      8000-32000

      Saliva (n=10)

      Healthy human

      30-613ng/mL

      2-50

      Urine (n=12)

      Healthy human

      27-138ng/mL

      2-10


      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human CLU were tested 20 times on one plate, respectively.

      Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human CLU 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)

      4.14

      11.58

      48.58

      4.26

      10.17

      47.33

      Standard deviation

      0.25

      0.57

      2.52

      0.24

      0.48

      2.06

      CV (%)

      6.04

      4.92

      5.19

      5.63

      4.72

      4.35



      Recovery

      The recovery of Human CLU 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=14)

      88-109

      97

      EDTA plasma (n=11)

      81-116

      92

      Salivan=10

      83-104

      91

      Urinen=12

      81-117

      91


      Linearity

      Samples were spiked with high concentrations of Human CLU 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)

      Saliva (n=4)

      Urinen=4

      1:2

      Range (%)

      81-114

      81-116

      87-120

      83-105

      Average (%)

      100

      99

      106

      93

      1:4

      Range (%)

      83-114

      82-117

      87-114

      81-99

      Average (%)

      98

      100

      100

      93

      1:8

      Range (%)

      89-117

      94-108

      86-115

      83-95

      Average (%)

      104

      100

      96

      91

      1:16

      Range (%)

      91-119

      91-116

      87-110

      88-99

      Average (%)

      105

      105

      99

      94


      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)

      8 wells ×12 strips

      2-8, 1 months

      Reference Standard

      2 vials

      Discard unused reconstituted standard and dilutions

      Reference Standard & Sample Diluent

      1 vial, 20 mL

      2-8

      Biotinylated Detection Ab Working Solution

      1 vial, 12 mL

      HRP Conjugate Diluent

      1 vial, 14 mL

      Concentrated Wash Buffer (25×)

      1 vial, 30 mL

      Concentrated HRP Conjugate (100×)

      1 vial, 120 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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