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MAK039

Sigma-Aldrich

Acetyl-Coenzyme A Assay Kit

sufficient for 100 fluorometric tests

Synonym(s):

Acetyl-CoA Assay Kit

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About This Item

UNSPSC Code:
12161503
NACRES:
NA.84

usage

sufficient for 100 fluorometric tests

detection method

fluorometric

relevant disease(s)

cancer; neurological disorders

storage temp.

−20°C

General description

Acetyl-CoA is an essential cofactor and carrier of acyl groups in enzymatic acetyl transfer reactions. It is formed either by the oxidative decarboxylation of pyruvate in mitochondria, by the oxidation of long-chain fatty acids, or by the oxidative degradation of certain amino acids. Acetyl-CoA is the starting compound for the citric acid cycle (Kreb′s cycle). It is also a key precursor in lipid biosynthesis, and the source of all fatty acid carbons. Acetyl-CoA positively regulates the activity of pyruvate carboxylase. It is a precursor of the neurotransmitter acetylcholine. Histone acetylases (HAT) use Acetyl-CoA as the donor for the acetyl group used in the post-translational acetylation reactions of histone and non-histone proteins.

Acetyl-Coenzyme A acts as an indicator of fat, sugar, and protein levels and shows up on the nutritional status. Thus, starvation greatly reduces the level of acetyl-CoA, which stimulates the process of autophagy.[1] In lowered glucose condition, acetyl-CoA participates in the production of ATP.[2] Increased acetyl-CoA levels are observed in prostate cancer due to elevated fatty acid utilization and thus provides an additional energy source for the tumor cell growth.[3] Nutrition deprivation, changes the levels of certain metabolites, including acetyl-CoA, which stimulates epigenetic modifications that would affect the central nervous system.[4]

Application

Acetyl-Coenzyme A Assay Kit has been used to quantify the level of acetyl-CoA in astrocytes exposed to recombinant HIV protein Tat and Cocaine.[5] It has also been used to study the mitochondrial metabolism by evaluating the levels of acetyl-CoA using hypoxia induced stress in primary human ventricular cardiomyocytes.[6]

Suitability

Suitable for the measurement of Acetyl CoA in a variety of biological samples

Principle

Acetyl-CoA concentration is determined by a coupled enzyme assay, which results in a fluorometric (λex = 535/λem = 587 nm) product, proportional to the Acetyl-CoA present. Typical sensitivities of detection for this kit are 10-1000 pmole of Acetyl CoA. This kit is a highly sensitive assay for determining Acetyl-CoA level in a variety of biological samples.

related product

Product No.
Description
Pricing

Pictograms

Health hazardExclamation mark
GHS08,GHS07

Signal Word

Danger

Hazard Statements

H315,H319,H334,H335

Hazard Classifications

Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3

Target Organs

Respiratory system

Storage Class Code

10 - Combustible liquids

Flash Point(F)

188.6 °F - closed cup

Flash Point(C)

87 °C - closed cup

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Certificates of Analysis (COA)

Lot/Batch Number
9A03K03170
9A05K03170
8A21K03170
8C24K03170
8D12K03170

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Epigenetic mechanisms in neurological and neurodegenerative diseases.
Jorge L G, et al.
Frontiers in Cellular Neuroscience, 9, 58-58 (2015)
François Gagné et al.
Comparative biochemistry and physiology. Toxicology & pharmacology : CBP, 172-173, 36-44 (2015-05-11)
The purpose of this study was to determine the cumulative effects of exposure to either dissolved zinc or nanozinc oxide (nanoZnO) and air-time survival in freshwater mussels. Mussels were exposed to each forms of zinc for 96h then placed in
Maria I Dauden et al.
Science advances, 5(7), eaaw2326-eaaw2326 (2019-07-17)
The highly conserved Elongator complex modifies transfer RNAs (tRNAs) in their wobble base position, thereby regulating protein synthesis and ensuring proteome stability. The precise mechanisms of tRNA recognition and its modification reaction remain elusive. Here, we show cryo-electron microscopy structures
Lilong He et al.
Plant physiology, 180(1), 198-211 (2019-02-17)
Cadmium (Cd) is a major heavy metal pollutant, and Cd toxicity is a serious cause of abiotic stress in the environment. Plants protect themselves against Cd stress through a variety of pathways. In a recent study, we found that mitochondrial
Nuclear-cytoplasmic localization of acetyl coenzyme a synthetase-1 in the rat brain
Ariyannur P S, et al.
The Journal of Comparative Neurology, 518(15), 2952-2977 (2010)
View All Related Papers

Articles

Glutamine Metabolism in Cancer Cells

Sigma article discusses tumor cell metabolic pathways, focusing on aerobic glycolysis and mitochondrial activity.

Aerobic Glycolysis and the Warburg Effect

Warburg effect enhances glucose to lactate conversion in tumor cells, regardless of oxygen levels; impacting cancer metabolism since 1924.

Fatty Acid Synthesis in Cancer Cells

Fatty acid synthesis supports cancer cell proliferation, essential for membrane generation, protein modification, and bioenergetics.

Questions

1–8 of 8 Questions  

  1. How is shipping temperature determined? And how is it related to the product storage temperature?

    1 answer

    1. 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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  2. How can I determine the shelf life / expiration / retest date of this product?

    1 answer

    1. 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/449/386/product-dating-information-mk.pdf

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  3. Can I use the Acetyl-CoA Assay Buffer to dissolve cells for the MAK039 Acetyl-Coenzyme A Assay Kit, even though the protocol suggests homogenizing the sample first?

    1 answer

    1. The following protocol is recommended for using the kit to assay cultured cells:

      Cell Lysis Buffer (1X):

      20 mM Tris (pH 7.5)
      150 mM NaCl
      1 mM EDTA
      1 mM EGTA
      1% Triton X-100
      2.5 mM sodium pyrophosphate
      1 mM B-Glycerolphosphate
      1 mM Na3VO4
      1 µg/ml Leupeptin
      1 mM PMSF before use.
      Preparing Cell Lysates:

      Aspirate media. Treat cells by adding fresh media containing a regulator for the desired time.
      To harvest cells under nondenaturing conditions, remove media and rinse cells once with ice-cold PBS.
      Remove PBS and add 0.5 ml 1X ice-cold Cell Lysis Buffer plus 1 mM PMSF to each plate (10 cm2) and incubate the plate on ice for 5 minutes.
      Scrape cells off the plate and transfer to microcentrifuge tubes. Keep on ice.
      Sonicate 4 times for 5 seconds each on ice.
      Microcentrifuge for 10 minutes at 4°C, and transfer the supernatant to a new tube. The supernatant is the cell lysate. If necessary, lysate can be stored at –80°C.
      Once the lysate is ready, follow the deproteinization protocol as mentioned on the attached datasheet and then proceed with the whole protocol.

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  4. I want to use cells from cell culture for this assay. 1) In what volume do I collect 2*10^6 cells prior to PCA precipitation? 2) Is PCA precipitation equal to lysis? 3) How do I verify pH with 1uL of sample? Thank you

    1 answer

    1. 1) In what volume do I collect 2*10^6 cells prior to PCA precipitation?
      Please count the cells with a hemocytometer and then harvest ~ 2 x 10^6 cells. There is not a specific volume that is collected as the end-user will have to understand the amount of cells/mL of cell suspension. Once the cells are harvested the end-user will resuspend the cell pellet in 500 uL of assay buffer on ice. This can be homogenized on ice for 10- 50 passes until the end-user can confirm lysis by viewing it under a microscope. The end-user will then do a centrifugation at 10,000 g for 10 minutes at 4C and collect the supernatant for PCA deproteinization.

      2) Is PCA precipitation equal to lysis?
      No. PCA precipitation is a deproteinization. Lysis should be completed by resuspending cells in an assay buffer with a dounce homogenizer.

      3) How do I verify pH with 1uL of the sample?
      Please use pH paper.

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  5. Where is the protocol for this kit?

    1 answer

    1. Please see the link below to review the product datasheet which includes the protocol:
      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/163/805/mak039bul.pdf

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  6. Can this kit be used for measuring Acetyl CoA levels in bacterial culture samples?

    1 answer

    1. Yes, this kit is a highly sensitive assay for determining Acetyl-CoA level in a variety of biological samples. See the link below to the technical bulletin:
      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/163/805/mak039bul.pdf

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  7. What is the Department of Transportation shipping information for this product?

    1 answer

    1. Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.

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  8. Can these kits be used with serum samples from mouse?

    1 answer

    1. Yes, this kit can be used on mouse serum samples. We generally recommend that fresh serum samples be used. Frozen samples can probably be used. However, the successful use of frozen samples depends on the efficiency of freezing them and the amount of time they have been frozen. If previously frozen serum samples are used, you may want to check the performance of the kit with fresh vs. frozen samples.

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