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MAK039

Sigma-Aldrich

乙酰辅酶A检测试剂盒

sufficient for 100 fluorometric tests

别名:

乙酰辅酶A检测试剂盒

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About This Item

UNSPSC代码:
12161503
NACRES:
NA.84

用途

sufficient for 100 fluorometric tests

检测方法

fluorometric

相关疾病

cancer; neurological disorders

储存温度

−20°C

一般描述

乙酰辅酶A是酶促的乙酰转移反应中酰基的必需辅助因子和载体。它可通过线粒体中丙酮酸的氧化脱羧、长链脂肪酸的氧化或某些特定氨基酸的氧化降解而形成。乙酰辅酶A是柠檬酸循环（克雷布氏循环）的起始化合物。它也是脂质生物合成的关键前体，所有脂肪酸碳的来源。乙酰辅酶A正向调节丙酮酸羧化酶活性。它是神经递质乙酰胆碱的前体。组蛋白乙酰基转移酶（HAT）在组蛋白和非组蛋白翻译后乙酰反应中使用乙酰辅酶A作为乙酰基供体。

乙酰辅酶A是脂肪，糖和蛋白质水平的指示剂，表明营养水平。因此，饥饿能够很大程度地降低乙酰辅酶A水平，激活自吞噬过程。[1]在低葡萄糖条件下，乙酰辅酶A参与ATP生成。[2]因为脂肪酸利用升高，在前列腺癌中观察到乙酰辅酶A水平升高，为肿瘤细胞生长提高额外能量。[3]营养不良会引起某些代谢物水平改变，包括乙酰辅酶A，激活表观遗传修饰，影响中枢神经系统。[4]

应用

乙酰辅酶A试剂盒可以用来定量暴露于重组HIV蛋白Tat和可卡因的胶细胞中乙酰辅酶A水平。[5]它还可以用于通过缺氧在原代人心室肌细胞引发压力，提高乙酰辅酶A水平，然后研究线粒体代谢的研究。[6]

适用性

适用于检测多种生物样品中的乙酰辅酶A。

原理

乙酰辅酶A浓度能够被偶联酶分析检测，生成的荧光 (λex = 535/λem = 587 nm)产物和乙酰辅酶A成正比。该试剂盒通常的检测灵敏度是10-1000pmole的乙酰辅酶A。该试剂盒检测各种生物样本中的乙酰辅酶A的灵敏度都很高。

相关产品

产品编号
说明
价格

象形图

Health hazardExclamation mark
GHS08,GHS07

警示用语：

Danger

危险分类

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

靶器官

Respiratory system

储存分类代码

10 - Combustible liquids

闪点(°F)

188.6 °F - closed cup

闪点(°C)

87 °C - closed cup

历史批次信息供参考:

分析证书(COA)

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

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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
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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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