15-Epi-Lipoxin A4 ELISA Assay


The 15-Epi-Lipoxin A4 ELISA Assay is for the quantitative determination of 15-Epi-Lipoxin A4 in biological fluids by a microplate enzyme immunoassay (ELISA).

SKU: 15E39-K01 Categories: , ,

15-Epi-Lipoxin A4 ELISA Assay

The 15-Epi-Lipoxin A4 ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 0.02 ng/ml
Dynamic Range: 0.02 – 20 ng/ml
Incubation Time: 2 hour
Sample Type: Biological Fluids
Sample Size: 1 mL

Product manufactured in the USA

Assay Principle

The 15-Epi-Lipoxin A4 ELISA Assay Kit (Enzyme-Linked Immunosorbent Assay) for the quantitative analysis of 15-epi-Lipoxin A4 levels in biological fluid. This test kit operates on the basis of competition between the enzyme conjugate and the 15-epi-LXA4 in the sample for a limited number of antibody binding sites.  The sample or standard solution is first 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 that 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-LXA4 in the sample or standard. For example, the absence of 15-epi-LXA4 in the sample will result in a bright blue color, whereas the presence of 15-epi-LXA4 will result in decreased or no color development.

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Additional Information

Assay Background

15-epi-Lipoxin A4 (15-epi-LXA4) 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.  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.

Assay Procedure

  1. Add 50 µL of Standards or Samples (may require diluting) to the corresponding wells on the microplate in duplicate.  See Scheme I for a sample plate layout.
  2. Add 50 µL of diluted 15-epi-LXA4-HRP Conjugate to each well. Incubate at room temperature for one hour.
  3. Wash the plate three times with 300 µL of diluted Wash Buffer per well. Wash 5 times if using an automated plate washer.
  4. Add 150 µL of TMB Substrate to each well. Incubate at room temperature for 30 minutes.
  5. Read the plate at 650 nm.

Alternately, the color reaction can be stopped after 10-15 minutes by adding 50 µL of 1 N HCl and read at 450 nm.

Cross Reactivity Data

15-epi-Lipoxin A4 100.0% 5(S)-HETE <0.01%
Lipoxin A4 3% 12(S)-HETE <0.01%
15(R)-HETE 0.8% 15(S)-HETE <0.01%

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