QuicKey Human Cys-C(Cystatin C) ELISA Kit

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

      Catalog number:E-TSEL-H0008

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

      Reactivity: Human

      Detection Range: 0.31~20 ng/mL

      Sensitivity: 0.04 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 Cys-C concentrations in serum, plasma, urine, saliva. Please consult technical support for the applicability if other biological fluids need to be tested.

       

      Background

      Cystatin C is a non-glycosylated cysteine protease inhibitor with a molecular weight of 13 kDa. It is produced at a constant rate by nucleated cell and freely filtered by the renal glomerulus, and serum cystatin C is an established, novel marker of renal function. Cystatin C is a more specific marker of glomerular filtration rate than creatinine, because levels are independent of age, sex, muscle mass and diet [1]. In addition, high cystatin C is associated with elevated risk of death from myocardial infarction, stroke, and metabolic syndrome. Higher levels of cystatin C and the presence of certain isoforms have also been related to neurological and cerebral disorders such as cerebral amyloid angiopathy, amyotrophic lateral sclerosis, multiple sclerosis, and Alzheimer’s disease. Cystatin isoforms can be detected both in serum and cerebrospinal fluid and more than a dozen have been reported so far.

       

      1. Ferguson T W, Komenda P, Tangri N. Cystatin C as a biomarker for estimating glomerular filtration rate. [J]. Curr Opin Nephrol Hypertens, 2015, 24(3):295-300.

       

      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 Cys-C. Samples (or Standards) and biotinylated detection antibody specific for Human Cys-C are added to the micro ELISA plate wells. Human Cys-C 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 Cys-C, 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 Cys-C. You can calculate the concentration of Human Cys-C 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

      0.31-20 ng/mL

      Sensitivity

      0.04 ng/mL

      Sample volume

      50μL

      Sample type

      Serum, Plasma, Urine, Saliva

       

      Specificity

      This kit recognizes Human Cys-C in samples. No significant cross-reactivity or interference between Cys-C 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.


      Typical standard curve when assay with the Reference Standard & Sample Diluent 1.

      Concentration(ng/mL)

      20

      10

      5

      2.5

      1.25

      0.63

      0.31

      0

      OD

      2.576

      1.625

      0.934

      0.574

      0.347

      0.209

      0.136

      0.069

      Corrected OD

      2.507

      1.556

      0.865

      0.505

      0.278

      0.14

      0.067

      -

       

       

      Typical standard curve when assay with the Reference Standard & Sample Diluent 2.

      Concentration(ng/mL)

      20

      10

      5

      2.5

      1.25

      0.63

      0.31

      0

      OD

      2.656

      2.183

      1.477

      0.904

      0.504

      0.283

      0.161

      0.059

      Corrected OD

      2.597

      2.124

      1.418

      0.845

      0.445

      0.224

      0.102

      -

      Sample values

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

      Sample Type

      Source

      Range

      Dilution Factor

      Serum (n=12)

      Healthy human

      442-1390ng/mL

      100-500

      EDTA plasma (n=13)

      Healthy human

      360-1295ng/mL

      100-500

      Saliva (n=13)

      Healthy human

      395-2948ng/mL

      100-800

      Urine (n=14)

      Healthy human

      4.3-58.2ng/mL

      1-20


      Precision

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

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

      0.86

      2.35

      9.01

      0.86

      2.06

      8.64

      Standard deviation

      0.05

      0.11

      0.37

      0.05

      0.1

      0.38

      CV (%)

      5.81

      4.68

      4.11

      5.81

      4.85

      4.4


      Recovery

      The recovery of Human Cys-C 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=12)

      90-101

      94

      EDTA plasma (n=13)

      96-108

      101

      Saliva(n=13)

      92-111

      105

      Urine(n=14)

      91-105

      99


      Linearity

      Samples were spiked with high concentrations of Human Cys-C 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 (%)

      95-109

      91-100

      83-108

      85-119

      Average (%)

      101

      95

      95

      102

      1:4

      Range (%)

      100-111

      82-93

      86-116

      103-118

      Average (%)

      105

      90

      105

      112

      1:8

      Range (%)

      86-98

      89-98

      80-113

      85-119

      Average (%)

      91

      92

      98

      105

      1:16

      Range (%)

      90-104

      91-101

      86-98

      96-107

      Average (%)

      96

      97

      92

      101


      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

      1 vial, 20 mL

      2-8℃

      Reference Standard & Sample Diluent 2

      1 vial, 20 mL

      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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