QuicKey Human IgM(Immunoglobulin M) 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-H0010

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

      Reactivity: Human

      Detection Range: 7.81~500 ng/mL

      Sensitivity: 1.02 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 IgM concentrations in serum, plasma, urine, saliva. Please consult technical support for the applicability if other biological fluids need to be tested.

       

      Background

      Immunoglobulin M (IgM) is a class of immunoglobulins found in blood and lymph fluid that are the first type of antibody made in response to an infection [1]. IgM antibodies make up about 5 percent to 10 percent of all the antibodies in the body. IgM is principally concerned with the primary antibody response, appearing soon after initial invasion by an antigen and capable of destroying the antigen when it is first introduced. Its normal mean of serum concentration is 120 mg/dl. IgM is the first immunoglobulin expressed in the human fetus (around 20 weeks) and phylogenetically the earliest antibody to develop [2]. IgM antibodies appear early in the course of an infection and usually reappear, to a lesser extent, after further exposure. IgM antibodies do not pass across the human placenta (only isotype IgG). The biological properties of IgM make it useful in the diagnosis of infectious diseases. Demonstrating IgM antibodies in a patient's serum indicates recent infection, or in a neonate's serum indicates intrauterine infection (e.g. congenital rubella syndrome).

       

      1. Gupta S, Pahwa R, O'Reilly R, et al. Ontogeny of lymphocyte subpopulations in human fetal liver. [J]. Proceedings of the National Academy of Sciences of the United States of America, 1976, 73(3):919-922.

      2. Van F R, Schuit H R, Hijmans W. The immunological development of the human fetus. [J]. Journal of Experimental Medicine, 1965, 122(6):1173.

       

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

      7.81-500 ng/mL

      Sensitivity

      1.02 ng/mL

      Sample volume

      50μL

      Sample type

      Serum, Plasma, Urine, Saliva

       

      Specificity

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

      500

      250

      125

      62.5

      31.25

      15.63

      7.81

      0

      OD

      2.594

      1.928

      1.405

      0.836

      0.524

      0.315

      0.188

      0.063

      Corrected OD

      2.531

      1.865

      1.342

      0.773

      0.461

      0.252

      0.125

      -

       


      Sample values

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

      Sample Type

      Source

      Range

      Dilution Factor

      Serum(n=14)

      Healthy human

      0.68-3.78mg/mL

      20000-80000

      EDTA plasma(n=14)

      Healthy human

      0.70-4.53mg/mL

      20000-80000

      Saliva(n=12)

      Healthy human

      6.4-35μg/mL

      50-500

      Urine(n=12)

      Healthy human

      ND

      -

       

      Precision

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

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

      20.4

      57.6

      229.31

      21.61

      62.32

      248.67

      Standard deviation

      1.21

      2.63

      12.59

      1.11

      3

      12.86

      CV (%)

      5.93

      4.57

      5.49

      5.14

      4.81

      5.17

       

      Recovery

      The recovery of Human IgM 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)

      93-101

      98

      EDTA plasma (n=14)

      97-106

      100

      Saliva(n=12)

      88-95

      91

      Urine(n=12)

      80-112

      104

       

      Linearity

      Samples were spiked with high concentrations of Human IgM 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)

      Urine (n=4)

      1:2

      Range (%)

      91-102

      84-98

      89-100

      92-120

      Average (%)

      98

      91

      97

      104

      1:4

      Range (%)

      94-106

      99-110

      91-102

      105-117

      Average (%)

      102

      102

      99

      110

      1:8

      Range (%)

      96-104

      87-94

      97-110

      88-115

      Average (%)

      99

      91

      104

      99

      1:16

      Range (%)

      88-98

      88-99

      102-112

      87-120

      Average (%)

      94

      92

      105

      106

       

      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

      37Incubator

      Deionized or distilled water

      Absorbent paper

      Loading slot for Wash Buffer


      SUMMARY

       


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