QuicKey Human TF(Transferrin) 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-H0001

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

      Reactivity: Human

      Detection Range: 0.14~100 ng/mL

      Sensitivity: 0.13 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 TF concentrations in serum, plasma, urine. Please consult technical support for the applicability if other biological fluids need to be tested.


      Background

      Transferrin is an iron binding protein found in blood plasma that is critical to cell health. Transferrin supplies iron to cells naturally in the body and as a supplement in cell culture media Transferrin (Tf) carries ferric iron from the intestine, reticuloendothelial system, and liver parenchymal cells to all proliferating cells in the body [1]. It shares homologous amino acid sequences with four other proteins: lactotransferrin, ovotransferrin, melanoma antigen p97, and HuBlym-1. Antigen p97 and the Tf receptor genes have been mapped on human chromosome 3. Transferrin has a molecular weight of around 80 kDa and contains two specific high-affinity Fe (III) binding sites. The affinity of transferrin for Fe (III) is extremely high (association constant is 1020M−1 at pH 7.4) but decreases progressively with decreasing pH below neutrality [2]. When not bound to iron, transferrin is known as Apo transferrin.


      1. Sousa M D , Macedo M F . Transferrin and the Transferrin Receptor: Of Magic Bullets and Other Concerns [J]. Inflammation & Allergy Drug Targets, 2008, 7(1): 41-52.

      2. Aisen P A , Leibman A , Zweier J L . Stoichiometric and site characteristics of the binding of iron to human transferrin [J]. Journal of Biological Chemistry, 1978, 253(6):1930-193.


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

      0.14-100 ng/mL

      Sensitivity

      0.13 ng/mL

      Sample volume

      50μL

      Sample type

      Serum, Plasma, Urine


      Specificity

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

      33.33

      11.11

      3.70

      1.23

      0.41

      0.14

      0

      OD

      2.647

      1.937

      1.181

      0.595

      0.297

      0.182

      0.123

      0.069

      Corrected OD

      2.578

      1.868

      1.112

      0.526

      0.228

      0.113

      0.054

      -



      Sample values

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

      Sample Type

      Source

      Range

      Dilution Factor

      Serumn=13

      Healthy human

      2.58-4.22mg/mL

      2×105-4×106

      EDTA plasman=13

      Healthy human

      2.13-4.42mg/mL

      2×105-4×106

      Urinen=11

      Healthy human

      51.45-1360.21ng/mL

      2-500

       

      Precision

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

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

      4.81

      12.27

      48.82

      4.88

      10.78

      48.95

      Standard deviation

      0.24

      0.6

      2.4

      0.26

      0.49

      2.36

      CV (%)

      4.99

      4.89

      4.92

      5.33

      4.55

      4.82

       

      Recovery

      The recovery of Human TF 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=13)

      87-94

      91

      EDTA plasma (n=13)

      90-99

      95

      Urine(n=11)

      95-107

      100

       

      Linearity

      Samples were spiked with high concentrations of Human TF 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)

      1:3

      Range (%)

      94-101

      92-102

      92-103

      Average (%)

      97

      95

      99

      1:9

      Range (%)

      97-106

      86-98

      98-111

      Average (%)

      102

      94

      105

      1:27

      Range (%)

      85-97

      94-107

      84-98

      Average (%)

      93

      101

      90

      1:81

      Range (%)

      101-109

      98-105

      85-99

      Average (%)

      104

      101

      91

       

      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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