Neogen ELISA Histamine Kit15-epi-Lipoxin A4 is an aspirin-triggered eicosanoid believed to be involved in the positive attributes of aspirin therapy for heart, cancer and human immunodeficiency virus patients. During inflammation, neutrophils are activated. 15-epi-LXA4, when administered in vivo, inhibits neutrophil activation and dampens inflammation (Takano, 1997). 15-epi-LXA4 is naturally formed in the body via this pathway:
Arachidonic Acid (AA)→15(R) -HETE→15-epi-LXA4

Aspirin is thought to be involved in the acetylation of prostaglandin G/H synthase, which triggers the conversion of AA to 15(R)-HETE. The development of this assay, which is both sensitive and selective for 15-epi-LXA4, will be essential to researchers studying anti-inflammatory medications.

This is an ELISA (Enzyme-Linked ImmunoSorbent Assay) for the quantitative analysis of 15-epi-Lipoxin A4 levels in biological fluid. This test kit operates based on competition between the enzyme conjugate and 15-epi-Lipoxin A4 in the sample for a limited number of binding sites.

First, the sample or standard solution is added to the microplate. Next, the diluted enzyme conjugate is added and the mixture is shaken and incubated at room temperature for one hour. During the incubation, competition for binding sites is taking place. The plate is then washed, removing all the unbound material. The bound enzyme conjugate is detected by the addition of substrate which generates an optimal color after 30 minutes. Quantitative test results may be obtained by measuring and comparing the absorbance reading of the wells of the samples against the standards with a microplate reader at 650 nm. The extent of color development is inversely proportional to the amount of 15-epi-Lipoxin A4 in the sample or standard. For example, the absence of 15-epi-Lipoxin A4 in the sample will result in a bright blue color, whereas the presence of 15-epi-Lipoxin A4 will result in decreased or no color development.

Standard Curve in EIA Buffer

Curve in EIA Buffer

Specificity (Cross-Reactivity)

Analytes Cross-Reactivity
15-epi-Lipoxin A4 100%
Lipoxin A4 3%
15(R)-HETE 0.8%
5(S)-HETE < 0.01%
12(S)-HETE < 0.01%
15(S)-HETE < 0.01%

Specifications

Sensitivity:
   80% B/B0 = 0.05 ng/mL
   50% B/B0 = 0.2 ng/mL
Assay Range: 0.2 - 2.0 ng/mL
Inter-Assay CV: ≤ 10%
Intra-Assay CV: ≤ 10%
Size: 96 well microplate
Assay Sample Size: 50 µL
Total Assay Incubation Time: 1.5 hours
Wavelength: 650 nm, 450 nm with acid stop
Storage Conditions: -20°C for lyophilized conjugate, 4°C for all other kit components
Antibody: Polyclonal, Rabbit

Order Details

Product # Product Description
407110 15-epi-Lipoxin A4, 96 well kit

Reference Materials

407110 - 15-epi-Lipoxin A4, Kit Insert [English]
407110 - 15-epi-Lipoxin A4, Safety Data Sheet [English]

Kit inserts available on our website are for reference use only and are subject to change. It is always recommended to use the protocol included with the kit.

Calculation of Results

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