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QuicKey Human TF(Transferrin) ELISA Kit

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Price:   $390
Cat Number:  E-TSEL-H0001
Lead Time:   7~10 days

Reactivity:  Human

Detection Range:  0.14~100 ng/mL

Sensitivity:  0.13 ng/mL

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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 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


SUMMARY


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