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LXA4(Lipoxin A4) ELISA Kit

Cat.No.:E-EL-0053
Reactivity: Universal

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96T
48T
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96T*5
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Product Details

Properties

Assay type	Competitive-ELISA
Format	96T/48T
Assay time	2.0h
Detection range	0.78-50 ng/mL
Sensitivity	0.47 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 50μL
Specificity	This kit recognizes LXA4 in samples. No significant cross-reactivity or interference between LXA4 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 LXA4 concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with LXA4. During the reaction, LXA4 in the sample or standard competes with a fixed amount of LXA4 on the solid phase supporter for sites on the Biotinylated Detection Ab specific to LXA4. Excess conjugate and unbound sample or standard are washed away, and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of LXA4 in tested samples can be calculated 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℃
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 LXA4 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level LXA4 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)	2.70	5.10	20.30	2.70	4.90	20.40
Standard deviation	0.10	0.20	1.00	0.10	0.20	0.80
CV (%)	3.70	3.92	4.93	3.70	4.08	3.92

Recovery

The recovery of LXA4 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)	92-106	100
EDTA plasma(n=8)	91-102	97
Cell culture media(n=8)	94-110	101

Linearity

Samples were spiked with high concentrations of LXA4 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Synonyms	ASPIRIN-TRIGGERED LXA4, LXA4-15-EPI, LIPOXIN A4, 15-EPI
Research Area	Cell Biology, Immunology

Assay Procedures

	1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 45 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

Citations

JOURNAL OF NANOBIOTECHNOLOGY (2022) IF: 9.429
Targeted delivery and ROS-responsive release of Resolvin D1 by platelet chimeric liposome ameliorates myocardial ischemia–reperfusion injury

DOI: 10.1186/s12951-022-01652-x

Sample: cardiac tissue,BMDMs
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY (2021) IF: 6.291
Pig lung fibrosis is active in the subacute CdCl2 exposure model and exerts cumulative toxicity through the M1/M2 imbalance

DOI: 10.1016/j.ecoenv.2021.112757

Sample: lung
Frontiers in Immunology (2018) IF: 5.511
Infective Larvae of Brugia malayi Induce Polarization of Host Macrophages that Helps in Immune Evasion

DOI: 10.3389/fimmu.2018.00194

Sample: Monocytes(MΦs)
JOURNAL OF CLINICAL PERIODONTOLOGY (2019) IF: 4.164
Salivary levels of specialized pro‐resolving lipid mediators as indicators of periodontal health/disease status

DOI: 10.1111/jcpe.13173

Sample: saliva
INTERNATIONAL IMMUNOPHARMACOLOGY (2022) IF: 4.932
LXA4 enhances prostate cancer progression by facilitating M2 macrophage polarization via inhibition of METTL3

DOI: 10.1016/j.intimp.2022.108586

PMID: 35228032
Sample: tumor tissue,pericarcinous tissue
INTERNATIONAL IMMUNOPHARMACOLOGY (2021) IF: 4.932
Lipoxin A4 activates ALX/FPR2 to attenuate inflammation in Aspergillus fumigatus keratitis

DOI: 10.1016/j.intimp.2021.107785

PMID: 34162149
Sample: Tissue homogenate

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