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AA(Arachidonic Acid) ELISA Kit

Cat.No.:E-EL-0051
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	1.56-100 ng/mL
Sensitivity	0.94 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 50μL
Specificity	This kit recognizes AA in samples. No significant cross-reactivity or interference between AA 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 AA 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 AA. During the reaction, AA in the sample or standard competes with a fixed amount of AA on the solid phase supporter for sites on the Biotinylated Detection Ab specific to AA. 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 AA 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 AA were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level AA 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)	4.70	9.10	36.70	4.70	8.70	36.60
Standard deviation	0.30	0.40	1.80	0.20	0.40	1.90
CV (%)	6.38	4.40	4.90	4.26	4.60	5.19

Recovery

The recovery of AA 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)	87-98	93
EDTA plasma(n=8)	93-110	101
Cell culture media(n=8)	90-103	98

Linearity

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

Target Information

Synonyms	ARA
Research Area	Signal transduction

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

Advanced Science (2022) IF: 17.521
Geometric Constraints Regulate Energy Metabolism and Cellular Contractility in Vascular Smooth Muscle Cells by Coordinating Mitochondrial DNA Methylation

DOI: 10.1002/advs.202203995

PMID: 36106364
Sample: Vascular smooth muscle cell (SMCs)
JCI Insight (2019) IF: 6.014
Systems biology identifies cytosolic PLA2 as a target in vascular calcification treatment

DOI: 10.1172/jci.insight.125638

Sample: human aortic smooth muscle cells (HAoSMCs)
Oncogenesis (2023) IF: 6.524
COX-2/PGE2 upregulation contributes to the chromosome 17p-deleted lymphoma

DOI: 10.1038/s41389-023-00451-9

Sample: lymphoma cell
Frontiers in Cell and Developmental Biology (2021) IF: 6.684
Fexofenadine Protects Against Intervertebral Disc Degeneration Through TNF Signaling

DOI: 10.3389/fcell.2021.687024

Sample: NP cell
Journal of Advanced Research (2019) IF: 5.045
Role of T lymphocytes and papain enzymatic activity in the protection induced by the cysteine protease against Schistosoma mansoni in mice

DOI: 10.1016/j.jare.2018.12.008

PMID: 31193307
Sample: Serum
Cell Death Discovery (2021) IF: 5.241
TRIB2 desensitizes ferroptosis via βTrCP-mediated TFRC ubiquitiantion in liver cancer cells

DOI: 10.1038/s41420-021-00574-1

PMID: 34315867
Sample: Bel-7404 cells,Bel-7402 cells,SMMC-7721 cells,SK-H
Frontiers in Endocrinology (2021) IF: 5.555
The Arachidonic Acid Metabolism Mechanism Based on UPLC-MS/MS Metabolomics in Recurrent Spontaneous Abortion Rats

DOI: 10.3389/fendo.2021.652807

Sample: plasma
Frontiers in Pharmacology (2022) IF: 5.811
Dapagliflozin Reduces Apoptosis of Diabetic Retina and Human Retinal Microvascular Endothelial Cells Through ERK1/2/cPLA2/AA/ROS Pathway Independent of Hypoglycemic.

DOI: 10.3389/fphar.2022.827896

Sample: Human retinal microvascular endothelial cells (HRM
Vaccines (2020) IF: 4.086
Specific Antibodies and Arachidonic Acid Mediate the Protection Induced by the Schistosoma mansoni Cysteine Peptidase-Based Vaccine in Mice

DOI: 10.3390/vaccines8040682

PMID: 33207535
Sample: Serum
Journal of Advanced Research (2018) IF: 4.327
Is arachidonic acid an endoschistosomicide?

DOI: 10.1016/j.jare.2018.01.005

Sample: serum
TOXICON (2016) IF: 2.309
Isolation and biochemical characterization of a gamma-type phospholipase A2 inhibitor from Macropisthodon rudis snake serum

DOI: 10.1016/j.toxicon.2016.09.011

Sample: Cell culture supernatant

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