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Human ApoA4(Apolipoprotein A4) ELISA Kit

Citations(6)Uniprot : P06727

Cat.No.:E-EL-H0463
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	4.69-300 ng/mL
Sensitivity	2.81 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human ApoA4 in samples. No significant cross-reactivity or interference between Human ApoA4 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 ApoA4 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 ApoA4. 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 ApoA4 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 ApoA4, 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 ApoA4. You can calculate the concentration of Human ApoA4 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 ApoA4 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human ApoA4 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)	14.00	45.80	115.80	12.70	47.40	118.00
Standard deviation	0.90	2.50	3.90	0.80	2.50	5.80
CV (%)	6.43	5.46	3.37	6.30	5.27	4.92

Recovery

The recovery of Human ApoA4 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)	90-102	95
EDTA plasma (n=8)	88-102	95
Cell culture media (n=8)	90-103	95

Linearity

Samples were spiked with high concentrations of Human ApoA4 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: P06727
Synonyms	APOA4,Apolipoprotein A-IV,ApoA-IV,ApoAIV,apoA4,
Research Area	Cancer, Cardiovascular, Metabolism, Signal transduction

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

ANALYTICA CHIMICA ACTA (2022) IF: 6.911
Investigating immunosensor for determination of depression marker-Apo-A4 based on patterning AuNPs and N-Gr nanomaterials onto ITO-PET flexible electrodes with amplifying signal

DOI: 10.1016/j.aca.2022.340217

PMID: 35998995
Sample: plasma
Scientific Reports (2016) IF: 5.228
iTRAQ-based proteomic analysis of plasma reveals abnormalities in lipid metabolism proteins in chronic kidney disease-related atherosclerosis

DOI: 10.1038/srep32511

Sample: Plasma
Frontiers in Neuroinformatics (2021) IF: 4.081
Bioinformatic Analysis of the Proteome in Exosomes Derived From Plasma: Exosomes Involved in Cholesterol Metabolism Process of Patients With Spinal Cord Injury in the Acute Phase

DOI: 10.3389/fninf.2021.662967

Sample: plasma
JOURNAL OF MOLECULAR AND CELLULAR CARDIOLOGY (2020) IF: 4.133
Identifying functional non-coding variants in APOA5/A4/C3/A1 gene cluster associated with coronary heart disease

DOI: 10.1016/j.yjmcc.2020.05.003

PMID: 32437778
Sample: Serum
BIOELECTROCHEMISTRY (2020) IF: 4.722
A novel electrochemical immunosensor for the highly sensitive and selective detection of the depression marker human apolipoprotein A4

DOI: 10.1016/j.bioelechem.2020.107542

PMID: 32388438
Sample: Plasma
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (2016) IF: 3.257
Label-Free Quantitative Proteomics Reveals Differences in Molecular Mechanism of Atherosclerosis Related and Non-Related to Chronic Kidney Disease

DOI: 10.3390/ijms17050631

Sample: Plasma

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