QuicKey Human IgM(Immunoglobulin M) ELISA Kit

    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
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    >
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience

      Catalog number:E-TSEL-H0010

      Size:
      • 96T
      • 48T
      Qty:
      - +
      Price: $390

      Reactivity: Human

      Detection Range: 7.80~500 ng/mL

      Sensitivity: 1.02 ng/mL

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

      Add to cart Compare Bulk request

      Product Details

      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.

      Properties

      Assay type Sandwich-ELISA
      Format 96T/48T
      Assay time 2.5 h
      Reactivity Human
      Detection method Colormetric
      Colormetric 7.80-500 ng/mL
      Sensitivity 1.02 ng/mL
      Sample volume 50μL/well
      Sample type serum, plasma, urine, saliva
      Specificity This kit recognizes Human IgM in samples. No significant cross-reactivity or interference between Human IgM and analogues was observed.
      Reproducibility Both intra-CV and inter-CV are < 10%.
      Application 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.

      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.

      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. (The specific components might be slightly different in delivered kits, please refer to the manual for more information.)

      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

      1. Microplate reader with 450nm wavelength filter

      2. High-precision transfer pipette, EP tubes and disposable pipette tips

      3. Incubator capable of maintaining 37℃

      4. Deionized or distilled water

      5. Absorbent paper

      6. Loading slot for Wash Buffer

      Technical Data

      Sample values

      sandwich-Ab-ELISA-Elabscience

      Typical Standard Curve

      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 31.3 15.63 7.80 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 -
      sandwich-Ab-ELISA-Elabscience

      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) 20.40 57.60 229.31 21.61 62.32 248.67
      Standard deviation 1.21 2.63 12.59 1.11 3.00 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=8) 93-101 98
      EDTA plasma(n=8) 97-106 100
      Urine(n=8) 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) Urine(n=4)
      1:2 Range (%) 91-102 84-98 92-120
      Average (%) 98 91 104
      1:4 Range (%) 94-106 99-110 105-117
      Average (%) 102 102 110
      1:8 Range (%) 96-104 87-94 88-115
      Average (%) 99 91 99
      1:16 Range (%) 88-98 88-99 87-120
      Average (%) 94 92 106

      Target information

      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, o
      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.

      Research area

      Documents

      Citations

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      Assay procedures

      step01 1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 90 min at 37°C
      step02 2. Aspirate and wash the plate for 3 times
      step03 3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
      step04 4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
      step05 5. Add 50μL Stop Solution
      step06 6. Read the plate at 450nm immediately. Calculation of the results
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      MSDS for ELISA ISO 9001

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