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Principaux documents

MAK363

Sigma-Aldrich

Lipoxygenase Activity Assay Kit

sufficient for 100 fluorometric tests

Synonyme(s) :

LOX Enzyme Activity Detection Kit

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About This Item

Code UNSPSC :
12352200

Utilisation

sufficient for 100 fluorometric tests

Méthode de détection

fluorometric

Description générale

Lipoxygenases (LOX) are non heme iron-containing dioxygenases. These enzymes convert unsaturated fatty acids to epoxides; for example, the synthesis of leukotrienes from arachidonic acid. They play an important role in cell proliferation, differentiation and inflammation; and have been implicated in inflammation and hyper proliferation mediated diseases like asthma, rheumatoid arthritis and cancer.

Adéquation

Suitable for the measurement of lipoxygenase activity in cell and tissue lysates, biological fluids (eg. serum), recombinant enzyme and purified protein.

Principe

In the Lipoxygenase Activity Assay Kit, lipoxygenase converts the LOX substrate to an intermediate that reacts with the probe generating a fluorescent product. The increase in fluorescent signal can be recorded at λex = 500 nm/λem = 536 nm and is directly proportional to LOX activity. The kit includes 5-lipoxygenase enzyme (LOX Enzyme) as a positive control. A lipoxygenase inhibitor that completely inhibits lipoxygenase activity is also included in order to calculate the specific activity of LOX in biological samples. The kit can detect as low as 0.004 mU/mg protein.

Pictogrammes

FlameHealth hazard

Mention d'avertissement

Danger

Mentions de danger

Classification des risques

Aquatic Chronic 3 - Eye Irrit. 2 - Flam. Liq. 2 - Resp. Sens. 1

Code de la classe de stockage

3 - Flammable liquids

Point d'éclair (°F)

49.5 °F

Point d'éclair (°C)

9.7 °C


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Questions

1–2 sur 2 questions  
  1. Could you provide the pH of the Assay buffer in the MAK363 Lipoxygenase Activity Assay Kit? Also, does the pH of the final solution interfere with the assay? If protein samples, extracted with high pH (10.8 buffer), are too diluted and need to add more samples, would the high pH of the samples inhibit the assay reaction?

    1 réponse
    1. The assay buffer has a pH of 7.4. It is recommended to maintain the sample pH closer to 7.4, as highly alkaline pH may interfere with the reaction.

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  2. What is the concentration of the enzyme?

    1 réponse
    1. The concentration of the enzyme standard for this kit is not reported. While it does confirm that the kit is functioning, it is not to be compared to samples or intended as a control. In order to quantify the LOX activity of the sample, please use the formula in step 6 on page 4 of the data sheet:
      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/247/994/mak363bul-ms.pdf

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