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QuicKey Human IgA(Immunoglobulin A) ELISA Kit

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Price:   $390
Cat Number:  E-TSEL-H0019
Lead Time:   7~10 days

Reactivity:  Human

Detection Range:  6.25~400 ng/mL

Sensitivity:  1.19 ng/mL

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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 6.25-400 ng/mL
Sensitivity 1.19 ng/mL
Sample volume 50μL/well
Sample type serum, plasma, urine, saliva
Specificity This kit recognizes Human IgA in samples. No significant cross-reactivity or interference between Human IgA 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 IgA 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 IgA. Samples (or Standards) and biotinylated detection antibody specific for Human IgA are added to the micro ELISA plate wells. Human IgA 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 IgA, 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 IgA. You can calculate the concentration of Human IgA 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) 400 200 100 50 25 62.5 6.25 0
OD 2.551 1.807 1.23 0.642 0.421 0.274 0.194 0.061
Corrected OD 2.491 1.746 1.169 0.582 0.36 0.213 0.133 -
sandwich-Ab-ELISA-Elabscience

Precision

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

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human IgA 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) 19.01 43.71 167.34 17.47 48.35 176.70
Standard deviation 1.09 2.12 8.32 0.88 2.40 9.33
CV (%) 5.73 4.85 4.97 5.04 4.96 5.28

Recovery

The recovery of Human IgA 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) 96-105 100
EDTA plasma(n=8) 98-106 101
Urine(n=8) 93-102 97

Linearity

Samples were spiked with high concentrations of Human IgA 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 (%) 88-103 86-97 100-112
Average (%) 95 93 104
1:4 Range (%) 97-110 94-103 93-102
Average (%) 105 99 97
1:8 Range (%) 94-109 99-111 85-95
Average (%) 101 103 92
1:16 Range (%) 93-105 91-104 90-98
Average (%) 97 98 93

Target information

Background

First described in serum in 1953, IgA is the second dominant isotype in the blood circulation following IgG. It can be found in both monomeric and polymeric forms. Circulating IgA is in monomeric form, whereas secretory IgA, in the mucosal secretions of respiratory, intestinal, and genitourinary systems, is dimeric. In humans, there are two subclasses of IgA: IgA1 and IgA2, constant heavy chains of which are encoded by two separate α1 and α2 genes on chromosome 14 [1]. The main structural difference between them is that IgA2 has a shorter hinge region which may render this isotype more resistant to bacterial proteases in the lumen of gastrointestinal or respiratory systems. In the blood, IgA interacts with an Fc receptor called FcαRⅠ(or CD89), which is expressed on immune effector cells, to initiate inflammatory reactions[2]. Ligation of FcαRⅠ by IgA containing immune complexes causes antibody-dependent cell-mediated cytotoxicity (ADCC), degranulation of eosinophils and basophils, phagocytosis by monocytes, macrophages, and neutrophils, and triggering of respiratory burst activity by polymorphonuclear leukocytes. IgA nephropathy is caused by IgA deposits in the kidneys, it is not yet known why IgA deposits occur in this chronic disease. Some theories suggest an abnormality of the immune system results in these deposits.
1. Kerr M A. Function of immunoglobulin A in immunity[J]. Gut, 2000, 47(6): 751-752.
2. Monteiro R C, Van De Winkel J G J. IgA Fc receptors[J]. Annual review of immunology, 2003, 21(1): 177-204.

Research area

Database Links

SwissProt Human
Entrez Gene Human

Synonyms

IgA

Documents

Citations

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Assay procedures

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
2. Aspirate and wash the plate for 3 times
3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
5. Add 50μL Stop Solution
6. Read the plate at 450nm immediately. Calculation of the results
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