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MAK005

Phenylalanine Assay Kit

sufficient for 100 fluorometric tests

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Informacje o tej pozycji

NACRES:
NA.84
UNSPSC Code:
12161503

usage

sufficient for 100 fluorometric tests

detection method

fluorometric

relevant disease(s)

neonatal diseases; neurological disorders; pediatric diseases

storage temp.

−20°C

General description

Phenylalanine is a non-polar essential amino acid. In the liver, phenylalanine is converted to tyrosine, which is a precursor for multiple compounds including L-DOPA and melanin. Defects in the activity of phenylalanine hydroxylase result in the inherited metabolic disorder phenylketonuria (PKU). PKU results in the build up of phenylalanine and phenylalanine metabolites, and can result in growth defects and mental retardation if not treated.

Application

Suitable for L-phenylalanine detection in cell and tissue culture supernatants, serum, and other biological samples

Biochem/physiol Actions

Phenylalanine concentration is determined by a coupled enzyme assay, which results in the deamination of phenylalanine and the production of NADH which reacts with the probe resulting in a fluorescent (λex = 535 nm/λem = 587 nm) product, proportional to the phenylalanine present.
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Piktogramy

Health hazard
GHS08

Słowo ostrzegawcze

Danger

Kody zagrożeń

H334

Ostrzeżenia

P261 - P284 - P501

Hazard Classifications

Resp. Sens. 1

Klasa składowania

10 - Combustible liquids

Temperatura zapłonu (°F)

No data available

Temperatura zapłonu (°C)

No data available

Co się dzieje?

WGK 3

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Certyfikaty analizy (CoA)

Lot/Batch Number

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Questions

1–2 of 2 Questions  

  1. What enzymes are included in the enzyme mix for this kit? Alternatively, please specify which enzymes are not part of the enzyme mix in the kit.

    1 answer

    1. The enzyme included in the kit is capable of reacting with tyrosine, methionine, and phenylalanine. According to a specific article, L-phenylalanine dehydrogenase can convert tyrosine and methionine in addition to phenylalanine.

      It's important to note that the enzyme is isolated from bacteria, not humans. The reaction pathway, likely catalyzed by phenylalanine dehydrogenase, is as follows: L-phenylalanine + H2O + NAD+ ⇌ phenylpyruvate + NH3 + NADH + H+.

      It was mentioned that any additional information about the enzyme would be proprietary. However, it was disclosed that this proprietary enzyme aids in the conversion of phenylalanine to generate NADH. In this process, PHE is reductively deaminated with the simultaneous formation of NADH, which reacts with our fluorescent probe to produce fluorescence at Ex/Em = 535/587 nm. It is advisable to proceed with the understanding that the reaction follows the pathway outlined in my previous note.

      Helpful?

  2. What is the procedure for removing the precipitate in the assay buffer?

    1 answer

    1. To dissolve the precipitate in the assay buffer, gently heat it with swirling. Placing it in a hot water bath at 50 - 60 degrees for 10-15 minutes should suffice. Avoid vigorously shaking the assay buffer to prevent foam formation. Allow it to cool to room temperature before proceeding with the assay.

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