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.
MAK052
ALT Activity Assay
sufficient for 100 colorimetric or fluorometric tests
Synonym(s):
Alanine Aminotransferase Activity Assay Kit, SGPT Activity Assay Kit, Serum Glutamic-Pyruvic Transaminase Activity Assay Kit
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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
Alanine Aminotransferase (ALT), also known as serum glutamic-pyruvic transaminase (SGPT), is a pyridoxal-phosphate-dependent enzyme that catalyzes the reversible transfer of an amino group from alanine to α-ketoglutarate, generating pyruvate and glutamate. ALT is found primarily in liver and serum, but occurs in other tissues as well. Hepatocellular injury often results in an increase of serum ALT levels and serum ALT levels can be used as a marker for liver injury.
Application
ALT Activity Assay kit has been used to determine the activity of alanine aminotransferase (ALT) in serum samples.[1][2][3]
Suitable for the measurement of alanine aminotransferase (ALT) activity in biological samples including tissue, cells and serum.
Biochem/physiol Actions
The ALT Activity Assay Kit provides a simple and direct procedure for measuring ALT activity in a variety of biological samples.
ALT activity is determined by a coupled enzyme assay, which results in a colorimetric (570 nm)/ fluorometric (λex = 535/λem = 587 nm) product, proportional to the pyruvate generated. One unit of ALT is defined as the amount of enzyme that generates 1.0 μmole of pyruvate per minute at 37 °C.
ALT activity is determined by a coupled enzyme assay, which results in a colorimetric (570 nm)/ fluorometric (λex = 535/λem = 587 nm) product, proportional to the pyruvate generated. One unit of ALT is defined as the amount of enzyme that generates 1.0 μmole of pyruvate per minute at 37 °C.
Features and Benefits
Compatible with high-throughput handling systems.
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Description
Pricing
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Danger
hcodes
Hazard Classifications
Aquatic Chronic 3 - Resp. Sens. 1 - Skin Sens. 1
Storage Class
10 - Combustible liquids
wgk_germany
WGK 3
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Instructions
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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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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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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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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
1 answer-
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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