QuicKey Human IgG(Immunoglobulin G) 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-H0011

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

      Reactivity: Human

      Detection Range: 1.56~100 ng/mL

      Sensitivity: 0.64 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 IgG concentrations in serum, plasma, urine, saliva. Please consult technical support for the applicability if other biological fluids need to be tested.

       

      Background

      Immunoglobulin G (IgG) is a class of immunoglobulins found in all body fluids. They are the smallest but most common antibodies (75 percent to 80 percent) in the body. IgG antibodies are large molecules of about 150 kDa [1], made of four peptide chains. It contains two identical class γ heavy chains of about 50 kDa and two identical light chains of about 25 kDa, thus a tetrameric quaternary structure. IgG antibodies are very important in fighting bacterial and viral infections, and they are the only type of antibody that can cross the placenta in a pregnant woman to help the fetus. The measurement of immunoglobulin G can be a diagnostic tool for certain conditions, such as autoimmune hepatitis, if indicated by certain symptoms. Clinically, measured IgG antibody levels are generally considered to be indicative of an individual's immune status to particular pathogens. A common example of this practice are titers drawn to demonstrate serologic immunity to measles, mumps, and rubella (MMR), hepatitis B virus, and varicella (chickenpox) [2].

       

      1. Jr C A J, Travers P, Walport M, et al. Principles of innate and adaptive immunity [J]. 2001.

      2. Faruque A A, Maoya J, et al. Analysis of surface plasmon resonance biosensors for protein detection. 2016.

       

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

      1.56-100 ng/mL

      Sensitivity

      0.64 ng/mL

      Sample volume

      50μL

      Sample type

      Serum, Plasma, Urine, Saliva

       

      Specificity

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

      2.532

      2.078

      1.476

      1.035

      0.635

      0.405

      0.245

      0.111

      Corrected OD

      2.421

      1.967

      1.365

      0.924

      0.524

      0.294

      0.134

      -

       


      Sample values

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

      Sample Type

      Source

      Range

      Dilution Factor

      Serum(n=12)

      Healthy human

      3.01-28.65mg/mL

      2×105-2×106

      EDTA plasma(n=12)

      Healthy human

      6.1-15.8mg/mL

      5×105-2×106

      Saliva(n=12)

      Healthy human

      ND-20846ng/mL

      1-2000

      Urine(n=12)

      Healthy human

      ND-1213ng/mL

      1-100

       

      Precision

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

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

      4.91

      11.2

      44.6

      4.94

      11.66

      47.68

      Standard deviation

      0.26

      0.51

      1.95

      0.25

      0.56

      2.24

      CV (%)

      5.3

      4.55

      4.37

      5.06

      4.8

      4.7

       

      Recovery

      The recovery of Human IgG 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)

      87-98

      93

      EDTA plasma (n=12)

      91-100

      95

      Saliva(n=12)

      85-94

      91

      Urine(n=12)

      83-107

      94

       

      Linearity

      Samples were spiked with high concentrations of Human IgG 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 (%)

      85-96

      87-97

      89-120

      86-94

      Average (%)

      90

      94

      105

      91

      1:4

      Range (%)

      89-98

      85-99

      92-116

      88-97

      Average (%)

      93

      92

      105

      91

      1:8

      Range (%)

      87-102

      92-102

      86-115

      94-107

      Average (%)

      94

      95

      103

      99

      1:16

      Range (%)

      86-98

      91-103

      86-123

      86-98

      Average (%)

      91

      97

      108

      90

       

      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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