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Mouse IFNα(Interferon Alpha) ELISA Kit

Citations(10)Uniprot : P01572,P01573

Cat.No.:E-EL-M3054
Reactivity: Mouse

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	15.63-1000 pg/mL
Sensitivity	9.38 pg/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Mouse IFNα in samples. No significant cross-reactivity or interference between Mouse 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 Mouse 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 Mouse 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 Mouse 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 Mouse 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 Mouse IFNα. You can calculate the concentration of Mouse 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 Mouse IFNα were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse 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)	49.31	114.27	450.06	52.89	117.30	463.36
Standard deviation	3.21	5.47	19.76	2.86	6.93	18.26
CV (%)	6.50	4.79	4.39	5.41	5.91	3.94

Recovery

The recovery of Mouse 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)	91-104	96
EDTA plasma (n=8)	92-108	99
Cell culture media (n=8)	90-104	97

Linearity

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

Target Information

Database Links	SwissProt: P01572 P01573
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

JOURNAL OF MEDICAL VIROLOGY (2022) IF: 20.693
FBXO6 regulates the anti-viral immune-responses via mediating alveolar macrophages survival

DOI: 10.1002/jmv.28203

PMID: 36217277
Sample: BALF
Acta Biomaterialia (2021) IF: 8.9469
Sialic acid conjugate-modified liposomes enable tumor homing of epirubicin via neutrophil/monocyte infiltration for tumor therapy

DOI: 10.1016/j.actbio.2021.07.063

PMID: 34339869
Sample: S180 cell-Xenograft
IMMUNOLOGY (2021) IF: 7.397
Neutralizing interferon-α blocks inflammation-mediated vascular injury via PI3K and AMPK in systemic lupus erythematosus

DOI: 10.1111/imm.13379

Sample: serum
Frontiers in Immunology (2021) IF: 7.561
Interplay Between Sialic Acids, Siglec-E, and Neu1 Regulates MyD88- and TRIF-Dependent Pathways for TLR4-Activation During Leishmania donovani Infection

DOI: 10.3389/fimmu.2021.626110

PMID: 33763070
Sample: Cell culture supernatant
EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY (2021) IF: 6.514
Antitumor activity of novel POLA1-HDAC11 dual inhibitors

DOI: 10.1016/j.ejmech.2021.113971

PMID: 34772529
Sample: Serum
Transboundary and Emerging Diseases (2020) IF: 4.188
H5N1 avian influenza virus without 80–84 amino acid deletion at the NS1 protein hijacks the innate immune system of dendritic cells for an enhanced mammalian pathogenicity

DOI: 10.1111/tbed.13904

Sample: Cell culture supernatant
ENDOCRINOLOGY (2021) IF: 4.736
Differential Effects of Viral Nucleic Acid Sensor Signaling Pathways on Testicular Sertoli and Leydig Cells

DOI: 10.1210/endocr/bqab180

PMID: 34453520
Sample: Leydig cell
Food & Function (2018) IF: 3.289
Structural characterization of two endopolysaccharides from Phellinus sp. and their immunologic effects by intragastric administration in a healthy mammalian model

DOI: 10.1039/C7FO01161J

Sample: serum
BMC IMMUNOLOGY (2022) IF: 3.594
NLRP3 inflammasome up-regulates major histocompatibility complex class I expression and promotes inflammatory infiltration in polymyositis

DOI: 10.1186/s12865-022-00515-2

PMID: 35965334
Sample: Raw 264.7 cell
JOURNAL OF FOOD BIOCHEMISTRY (2022) IF: 2.72
The immune-enhancing effect and in vitro antioxidant ability of different fractions separated from Colla corii asini

DOI: 10.1111/jfbc.14174

PMID: 35415887

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