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Human IFNα(Interferon Alpha ) ELISA Kit

Citations(8)Uniprot : P01562,P01563

Cat.No.:E-EL-H6125
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	7.81-500 pg/mL
Sensitivity	4.69 pg/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human IFNα in samples. No significant cross-reactivity or interference between Human IFNα 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 IFNα concentrations in serum, plasma and other biological fluids.

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 IFNα. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human IFNα and 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 IFNα, 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 nm ± 2 nm. The OD value is proportional to the concentration of Human IFNα. You can calculate the concentration of Human IFNα 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 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Substrate Reagent	1 vial, 10 mL	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8°C
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human IFNα were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human IFNα 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 (pg/mL)	23.29	65.91	237.77	21.20	60.10	221.45
Standard deviation	1.49	3.41	10.94	1.19	3.05	11.03
CV (%)	6.39	5.17	4.60	5.60	5.07	4.98

Recovery

The recovery of Human IFNα 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)	91-103	97
EDTA plasma (n=8)	87-98	92
Cell culture media (n=8)	91-105	98

Linearity

Samples were spiked with high concentrations of Human IFNα and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: P01562 P01563
Research Area	Immunology, Cancer

Assay Procedures

	1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
	2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
	3. Aspirate and wash the plate for 3 times
	4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	6. Add 50μL Stop Solution
	7. Read the plate at 450nm immediately. Calculation of the results

Citations

Signal Transduction and Targeted Therapy (2021) IF: 18.187

Differential immune responses in pregnant patients recovered from COVID-19

DOI: 10.1038/s41392-021-00703-3
PMID: 34326311	Sample: blood

Theranostics (2021) IF: 11.556

A SCID mouse-human lung xenograft model of SARS-CoV-2 infection

DOI: 10.7150/thno.58321
PMID: 33995679	Sample: Tissue homogenate

Frontiers in Immunology (2018) IF: 5.511

Potential Involvement of Platelet-Derived Microparticles and Microparticles Forming Immune Complexes during Monocyte Activation in Patients with Systemic Lupus Erythematosus

DOI: 10.3389/fimmu.2018.00322
	Sample: monocytes

INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (2021) IF: 5.924

2-O-Methylhonokiol Suppresses HCV Replication via TRAF6-Mediated NF-kB Activation

DOI: 10.3390/ijms22126499
PMID: 34204438	Sample: Huh7.5 cells

Journal of Inflammation Research (2022) IF: 4.631

miR-31-5p Regulates Type I Interferon by Targeting SLC15A4 in Plasmacytoid Dendritic Cells of Systemic Lupus Erythematosus

DOI: 10.2147/JIR.S383623
	Sample: serum

JOURNAL OF INTERFERON AND CYTOKINE RESEARCH (2022) IF: 3.657

Can the Prognosis of COVID-19 Disease Be Determined by Fecal Markers and Cytokines?

DOI: 10.1089/jir.2022.0098
PMID: 36179036	Sample: fecal

Innate Immunity (2019) IF: 2.173

Functional characterisation of Holothuria leucospilota Fas-associated death domain in the innate immune-related signalling pathways:

DOI: 10.1177/1753425919877680
	Sample: Cell culture supernatant

IMMUNOPHARMACOLOGY AND IMMUNOTOXICOLOGY (2020) IF: 2.352

Anti-inflammatory effects of nobiletin on TLR4/TRIF/IRF3 and TLR9/IRF7 signaling pathways in prostate cancer cells

DOI: 10.1080/08923973.2020.1725040
PMID: 32048561	Sample: Cell culture supernatant

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