QuicKey Human BMG/β2-MG(Beta-2-Microglobulin) ELISA Kit

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

      Catalog number:E-TSEL-H0004

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

      Reactivity: Human

      Detection Range: 0.78~50 ng/mL

      Sensitivity: 0.47 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 BMG/β2-MG concentrations in serum, plasma, urine. Please consult technical support for the applicability if other biological fluids need to be tested.



      Background

      Beta 2 microglobulin (β2M) is a small protein (11,800 Dalton), normally found on the surface of many cells, including lymphocytes, and in small amounts in the blood and urine [1]. An increased amount in the blood or urine may be a sign of certain diseases, including some types of cancer, such as multiple myeloma or lymphoma.β2M is critical for the immune surveillance and modulation in vertebrate animals. The dysregulation of β2M is associated with multiple diseases, including endogenous and infectious diseases. β2M could directly participate in the development of cancer cells, and the level of β2M is deemed as a prognostic marker for several malignancies. It also involves in forming major histocompatibility complex (MHC class I or MHC I) or like heterodimers, covering from antigen presentation to immune homeostasis [2].


      1. Güssow D, Rein R, Ginjaar I, et al. The human beta 2-microglobulin gene. Primary structure and definition of the transcriptional unit. [J]. Journal of Immunology, 1987, 139(9):3132-8.

      2. Ohta Y, Shiina T, Lohr R L, et al. Primordial linkage of β2-microglobulin to the MHC [J]. Journal of Immunology, 2011, 186(6):3563-3571.


      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 BMG/β2-MG. Samples (or Standards) and biotinylated detection antibody specific for Human BMG/β2-MG are added to the micro ELISA plate wells. Human BMG/β2-MG 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 BMG/β2-MG, 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 BMG/β2-MG. You can calculate the concentration of Human BMG/β2-MG 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

      0.18-50 ng/mL

      Sensitivity

      0.47 ng/mL

      Sample volume

      100μL

      Sample type

      Serum, Plasma, Urine


      Specificity

      This kit recognizes Human BMG/β2-MG in samples. No significant cross-reactivity or interference between BMG/β2-MG 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)

      50

      25

      12.5

      6.25

      3.13

      1.56

      0.78

      0

      OD

      2.744

      2.393

      1.756

      0.966

      0.483

      0.234

      0.149

      0.063

      Corrected OD

      2.681

      2.330

      1.693

      0.903

      0.420

      0.171

      0.086

      -




      Sample values

      Serum/Plasma/Urine–Samples from apparently healthy volunteers were evaluated for the presence of Human BMG/β2-MG in this assay.

      Sample Type

      Source

      Range

      Dilution Factor

      Serumn=12

      Healthy human

      1.4-2.7μg/mL

      100-2000

      EDTA plasman=12

      Healthy human

      1.1-2.4μg/mL

      100-2000

      Urinen=11

      Healthy human

      12.7-107ng/mL

      1-50


      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human BMG/β2-MG were tested 20 times on one plate, respectively.

      Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human BMG/β2-MG 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)

      2.12

      5.33

      23.47

      2.02

      5.9

      24.97

      Standard deviation

      0.12

      0.25

      1.06

      0.12

      0.28

      1.09

      CV (%)

      5.66

      4.69

      4.52

      5.94

      4.75

      4.37



      Recovery

      The recovery of Human BMG/β2-MG 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=11)

      84-104

      89

      EDTA plasma (n=11)

      81-118

      87

      Urinen=11

      76-123

      96


      Linearity

      Samples were spiked with high concentrations of Human BMG/β2-MG 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 (%)

      89-119

      88-94

      83-102

      Average (%)

      102

      91

      93

      1:4

      Range (%)

      92-120

      87-97

      94-105

      Average (%)

      106

      92

      100

      1:8

      Range (%)

      89-119

      92-104

      85-114

      Average (%)

      103

      101

      106

      1:16

      Range (%)

      89-103

      94-100

      93-105

      Average (%)

      95

      97

      99


      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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