QuicKey Human IgG(Immunoglobulin G) 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-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.

      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 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 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 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

      Assay time

      2.5h

      Reactivity

      Human

      Detection method

      Colormetric

      Detection range

      1.56-100 ng/mL

      Sensitivity

      0.64 ng/mL

      Sample volume

      100μL

      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

      2.716

      2.131

      1.540

      0.990

      0.597

      0.361

      0.246

      0.118

      Corrected OD

      2.598

      2.013

      1.422

      0.872

      0.479

      0.243

      0.128

      -



      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

      Serumn=12

      Healthy human

      2.99-28.02mg/mL

      2×105-2×106

      EDTA plasman=12

      Healthy human

      6.4-15.3mg/mL

      5×105-2×106

      Salivan=12

      Healthy human

      ND-20807ng/mL

      1-2000

      Urinen=12

      Healthy human

      ND-1138ng/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)

      4.55

      10.18

      45.63

      4.6

      12.24

      45.04

      Standard deviation

      0.23

      0.47

      2.48

      0.26

      0.58

      2.39

      CV (%)

      5.05

      4.62

      5.44

      5.65

      4.74

      5.31



      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)

      87-114

      103

      EDTA plasma (n=12)

      86-107

      97

      Salivan=12

      76-103

      85

      Urinen=12

      75-122

      90


      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 (%)

      79-97

      87-128

      90-123

      80-118

      Average (%)

      87

      106

      109

      107

      1:4

      Range (%)

      91-111

      81-94

      94-113

      97-127

      Average (%)

      99

      87

      103

      110

      1:8

      Range (%)

      99-118

      82-96

      84-117

      107-115

      Average (%)

      104

      90

      105

      112

      1:16

      Range (%)

      106-118

      85-114

      90-125

      107-115

      Average (%)

      112

      100

      110

      112


      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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