Data concerning the total incubation time during which a linear reaction rate would be maintained for samples with very high AST activity is not available. The positive control that was supplied with the now discontinued kit maintained a linear reaction rate through 60 minutes after an initial lag phase of approximately 10 minutes.
Fortfahren mit
MAK052
ALT-Aktivität Assay-Kit
sufficient for 100 colorimetric or fluorometric tests
Synonym(e):
Alanin-Aminotransferase-Aktivität Assay-Kit, SGPT-Aktivität Assay-Kit, Serum-Glutamat-Pyruvat-Transaminase-Aktivität Assay-Kit
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Über diesen Artikel
NACRES:
NA.84
UNSPSC Code:
12161503
usage
sufficient for 100 colorimetric or fluorometric tests
detection method
colorimetric, fluorometric
relevant disease(s)
gastrointestinal diseases
shipped in
wet ice
storage temp.
−20°C
Gene Information
human ... GPT(2875)
mouse ... GPT(76282)
rat ... GPT(81670)
General description
Alaninaminotransferase (ALT), auch Serum-Glutamat-Pyruvat-Transaminase (SGPT) genannt, ist ein Pyridoxalphosphat-abhängiges Enzym, das die reversible Transferase einer Aminogruppe von Alanin zu α-Ketoglutarat katalysiert und dadurch Pyruvat und Glutamat generiert. ALT tritt vor allem in der Leber und im Serum auf, kann aber auch in anderem Gewebe vorkommen. Eine hepatozelluläre Verletzung führt oft zu einem Anstieg der ALT-Konzentration im Serum, weshalb ALT-Konzentrationen als Marker für Leberverletzungen verwendet werden können.
Application
Das ALT-Aktivität Assay-Kit wurde für die Bestimmung der Aktivität von Alaninaminotransferase (ALT) in Serumproben verwendet.
Geeignet für die Messung der Alaninaminotransferase (ALT)-Aktivität in biologischen Proben, wie Gewebe, Zellen und Serum.
Biochem/physiol Actions
Das ALT-Aktivität Assay-Kit ermöglicht ein einfaches, direktes Verfahren für die Messung der ALT-Aktivität in verschiedenen biologischen Proben.
Die ALT-Aktivität wird durch einen gekoppelten Enzym-Assay bestimmt, der ein kolometrisches (570 nm)/ fluorometrisches (λex = 535/λem = 587 nm) Produkt ergibt, das proportional zum Pyruvat generiert wird. Eine ALT-Einheit ist definiert als die Menge an Enzym, die 1,0 μmol Pyruvat pro Minute bei 37 °C generiert.
Die ALT-Aktivität wird durch einen gekoppelten Enzym-Assay bestimmt, der ein kolometrisches (570 nm)/ fluorometrisches (λex = 535/λem = 587 nm) Produkt ergibt, das proportional zum Pyruvat generiert wird. Eine ALT-Einheit ist definiert als die Menge an Enzym, die 1,0 μmol Pyruvat pro Minute bei 37 °C generiert.
Features and Benefits
Kompatibel mit Handhabungssystemen mit hohem Durchsatz.
Signalwort
Danger
Gefahrencodes
Sicherheitshinweise
Hazard Classifications
Aquatic Chronic 3 - Resp. Sens. 1 - Skin Sens. 1
Lagerklasse
10 - Combustible liquids
Was ist los?
WGK 3
Hier finden Sie alle aktuellen Versionen:
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Could you tell me the average number of readings or typical total incubation time (at 37 °C) until the most active sample reaches the linear range limit?
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How can I determine the shelf life / expiration / retest date of this product?
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If this product has an expiration or retest date, it will be shown on the Certificate of Analysis (COA, CofA). If there is no retest or expiration date listed on the product's COA, we do not have suitable stability data to determine a shelf life. For these products, the only date on the COA will be the release date; a retest, expiration, or use-by-date will not be displayed.
For all products, we recommend handling per defined conditions as printed in our product literature and website product descriptions. We recommend that products should be routinely inspected by customers to ensure they perform as expected.
For products without retest or expiration dates, our standard warranty of 1 year from the date of shipment is applicable.
For more information, please refer to the Product Dating Information document: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/documents/418/501/product-dating-information-06-25-mk.pdfHelpful?
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Hi, Does this ALT kit depend on LDH for the ultimate readout? Our goal is to measure liver dysfunction in serum of mice. Would differences in serum LDH influence the results? Thanks Ricky
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This kit does not use lactate dehydrogenase (LDH) itself, but the measured results are proportional to the concentration of pyruvate generated. Because LDH (in the presence of NADH) can influence the concentration of pyruvate present, the presence of LDH in a sample may influence the assay results.
LDH was not assessed in the development of this kit, but the kit has been used to test serum samples directly. It is recommended to perform several sample dilutions to determine the optimal sample concentration and ensure measured values fall within the linear range of the standard curve. Liver cell and liver-derived cell line lysates are more commonly measured, as serum levels of ALT are normally low. Regardless, considering the effect of LDH on pyruvate in the presence of NADH, a variable concentration of LDH and/or NADH may impact the ALT assay results.
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How is shipping temperature determined? And how is it related to the product storage temperature?
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Products may be shipped at a different temperature than the recommended long-term storage temperature. If the product quality is sensitive to short-term exposure to conditions other than the recommended long-term storage, it will be shipped on wet or dry-ice. If the product quality is NOT affected by short-term exposure to conditions other than the recommended long-term storage, it will be shipped at ambient temperature. As shipping routes are configured for minimum transit times, shipping at ambient temperature helps control shipping costs for our customers. For more information, please refer to the Storage and Transport Conditions document: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/documents/316/622/storage-transport-conditions-mk.pdf
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Can I know the specific activity of this ALT mix sample? In unit of U/mg
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The ALT Enzyme Mix included in this kit is a lyophilized mixture of enzymes. The components include Pyruvate Oxidase, Cocarboxylase, and Sucrose. The concentration and activity of each enzyme are considered proprietary. Please see the link below to review the product datasheet:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/731/802/mak052bul.pdfHelpful?
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How does the presence of pyruvate in the Krebs buffer affect the final reaction of the kit, and why is its removal recommended? Additionally, what is the significance of the absence of enzymes in the kit for converting glucose and lactate to pyruvate in relation to the buffer?
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The kit's final reaction is dependent on the oxidation of pyruvate, so its presence in the buffer may interfere with the assay. If possible, it is recommended to remove pyruvate from the Krebs buffer. However, the kit does not contain enzymes that can convert glucose to pyruvate or lactate to pyruvate, so the presence of these should not pose an issue in the buffer.
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Is the ALT activity assay kit MAK052 compatible with EDTA plasma? Does EDTA interfere with the activity of the assay?
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EDTA has the potential to deactivate the enzyme. Therefore, it is recommended not to use samples containing EDTA for performing the AST and ALT assay kits, including the MAK055 kit.
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Can I use the kit for both mouse and human serum?
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This kit is suitable for mouse and human serum. As mentioned on the Product Detail Page, this item is suitable for measuring alanine aminotransferase (ALT) activity in biological samples including tissue, cells, and serum.
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Can it be used for measuring on supernatant from liver cells
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The kit is optimized for use in determination of alanine aminotransferase in biological samples (e.g. plasma, serum, cell lysate, tissue samples.) This kit has not been tested for use with cell culture supernatant. The presence of phenol red may interfere with results. In addition, the cultured cell line may not secrete sufficient ALT to be detected. It would be up to the end user to determine suitability.
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