Testing Data
15-Epi-Lipoxin A4 ELISA Kit
15-Epi-Lipoxin A4 EIA Kit
Synonyms
15-Epi-Lipoxin A4; 15-Epi-Lipoxin A4 EIA Kit; 15-epi-Lipoxin A4 Enzyme Immunoassay; 15-Epi-Lipoxin A4 elisa kit
Procedural Notes
1. It is not necessary to allow reagents to warm to room temperature before use.
2. Desiccant bag must remain in foil pouch with unused strips. Keep pouch sealed when not in use to maintain a dry environment. Seal with a heat sealer. If a heat sealer is not available, thoroughly close the open end with tape. Remove excess air before sealing.
3. Always use clean pipette tips for the buffer, enzyme conjugate, standards and samples.
4. Before pipetting a reagent, rinse the pipette tip three times with that reagent (i.e. fill the tip with the desired amount of reagent and dispense back into the same vial - repeat 2 times). Now the tip is properly rinsed and ready to dispense the reagent into your well or test tube.
5. When pipetting into the wells, DO NOT allow the pipette tip to touch the inside of the well, or any of the reagents already in the well. This can result in cross contamination.
6. Standards and samples should be assayed in duplicate.
7. To quantitate, always run a standard curve in duplicate when testing samples. If testing a sample that is not extracted, dilute standards in the same type of medium being tested. This medium should be known to be negative.
8. Gently mix specimens and reagents before use. Avoid vigorous agitation.
9. Before opening the lyophilized conjugate vial, examine the vial to ensure that lyophilized material has not been trapped in the cap. If material is in the cap, gently tap the upright vial to dislodge the trapped material.
10. To reconstitute the lyophilized conjugate, add 75 muL of deionized water to a vial. Rehydrate the conjugate by gently rotating the vial. Do not vortex or shake the contents. Avoid excess foaming. After the solid material has gone into solution, allow the conjugate to incubate at least 15 minutes before dilution. Write the date of reconstitution on the label. Concentrated, reconstituted conjugate has a shelf life of at least two weeks when stored at 4 degree C.
11. The enzyme conjugate is most stable in its concentrated form. Dilute only the volume necessary for the amount of strips currently being used. 1
2. Before taking an absorbance reading, wipe the outside bottom of the wells with a lintfree wiper to remove dust and fingerprints.
2. Desiccant bag must remain in foil pouch with unused strips. Keep pouch sealed when not in use to maintain a dry environment. Seal with a heat sealer. If a heat sealer is not available, thoroughly close the open end with tape. Remove excess air before sealing.
3. Always use clean pipette tips for the buffer, enzyme conjugate, standards and samples.
4. Before pipetting a reagent, rinse the pipette tip three times with that reagent (i.e. fill the tip with the desired amount of reagent and dispense back into the same vial - repeat 2 times). Now the tip is properly rinsed and ready to dispense the reagent into your well or test tube.
5. When pipetting into the wells, DO NOT allow the pipette tip to touch the inside of the well, or any of the reagents already in the well. This can result in cross contamination.
6. Standards and samples should be assayed in duplicate.
7. To quantitate, always run a standard curve in duplicate when testing samples. If testing a sample that is not extracted, dilute standards in the same type of medium being tested. This medium should be known to be negative.
8. Gently mix specimens and reagents before use. Avoid vigorous agitation.
9. Before opening the lyophilized conjugate vial, examine the vial to ensure that lyophilized material has not been trapped in the cap. If material is in the cap, gently tap the upright vial to dislodge the trapped material.
10. To reconstitute the lyophilized conjugate, add 75 muL of deionized water to a vial. Rehydrate the conjugate by gently rotating the vial. Do not vortex or shake the contents. Avoid excess foaming. After the solid material has gone into solution, allow the conjugate to incubate at least 15 minutes before dilution. Write the date of reconstitution on the label. Concentrated, reconstituted conjugate has a shelf life of at least two weeks when stored at 4 degree C.
11. The enzyme conjugate is most stable in its concentrated form. Dilute only the volume necessary for the amount of strips currently being used. 1
2. Before taking an absorbance reading, wipe the outside bottom of the wells with a lintfree wiper to remove dust and fingerprints.
Sample Preparation
Usually, urine and tissue culture supernatant can be assayed directly by diluting them with the diluted extraction buffer. Dilute specimens may require extraction in order to concentrate 15-epi-LXA
4. Plasma and most other mediums will need to be extracted.
4. Plasma and most other mediums will need to be extracted.
Preparation and Storage
4/ -20 degree C
Testing Data
Testing Data
Testing Data #2
Testing Data #2
Related Product Information for 15-Epi-Lipoxin A4 elisa kit
Description: 15-epi-Lipoxin A4 is an aspirin-triggered eicosanoid believed to be involved in the positive attributes of aspirin therapy for heart, cancer, and human immunodefiency 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.
Introduction: Adenosine 3':5'-cyclic monophosphate (cyclic AMP, cAMP) is one of the most important secondary messengers involved in signal transduction, gene regulation, steroid hormone biosynthesis and intracellular signaling. Its regulatory role extends to the nervous system, cell growth and differentiation, immune mechanisms and general metabolism. Cyclic AMP production is stimulated by compounds (hormones, drugs, neuromodulators) binding to target cells through cell membrane receptors. These binding events activate Gproteins which in turn activate the enzyme adenylate cyclase. Adenylate cyclase converts ATP to cyclic AMP. Cyclic AMP-dependent protein kinases can then be activated resulting in phosphorylation of substrate proteins. Cyclic AMP levels may aid in understanding the mechanisms of action of a variety of hormones and biologically active substances.
Principle of assay: 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 on the basis of 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.
Introduction: Adenosine 3':5'-cyclic monophosphate (cyclic AMP, cAMP) is one of the most important secondary messengers involved in signal transduction, gene regulation, steroid hormone biosynthesis and intracellular signaling. Its regulatory role extends to the nervous system, cell growth and differentiation, immune mechanisms and general metabolism. Cyclic AMP production is stimulated by compounds (hormones, drugs, neuromodulators) binding to target cells through cell membrane receptors. These binding events activate Gproteins which in turn activate the enzyme adenylate cyclase. Adenylate cyclase converts ATP to cyclic AMP. Cyclic AMP-dependent protein kinases can then be activated resulting in phosphorylation of substrate proteins. Cyclic AMP levels may aid in understanding the mechanisms of action of a variety of hormones and biologically active substances.
Principle of assay: 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 on the basis of 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.
References
1. Takano T, Fiore S, Maddox JF, Brady HR, Petasis NA, Serhan CN. Aspirin triggered 15-epi-Lipoxin A4 (LXA4) and LXA4 stable analogues are potent inhibitors of acute inflammation: evidence for anti-inflammatory receptors. J. Exp. Med. 185(9): 1693-1704 (1997).
2. Takano T, Clish CB, Gronert K, Petasis N, Serhan CN. Neutrophil-mediated changes in vascular permeability are inhibited by topical applications of aspirintriggered 15-epi-Lipoxin A4 and novel Lipoxin B4 stable analogues. J. Clin. Invest. 101: 819-826 (1998).
2. Takano T, Clish CB, Gronert K, Petasis N, Serhan CN. Neutrophil-mediated changes in vascular permeability are inhibited by topical applications of aspirintriggered 15-epi-Lipoxin A4 and novel Lipoxin B4 stable analogues. J. Clin. Invest. 101: 819-826 (1998).
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Product Notes
The 15-Epi-Lipoxin A4 (Catalog #AAA480522) is an ELISA Kit and is intended for research purposes only. The product is available for immediate purchase. It is sometimes possible for the material contained within the vial of "15-Epi-Lipoxin A4, ELISA Kit" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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