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MAK135

Sigma-Aldrich

ADP/ATP Ratio Assay Kit

sufficient for 100 tests (bioluminescent)

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1 KIT
647,00 $

647,00 $

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1 KIT
647,00 $

About This Item

Code UNSPSC :
12161503
Nomenclature NACRES :
NA.84

647,00 $

Veuillez contacter notre Service Clients pour connaître la disponibilité de ce produit.
Devis pour commande en gros

Utilisation

sufficient for 100 tests (bioluminescent)

Application(s)

pharmaceutical

Méthode de détection

chemiluminescent

Maladie(s) pertinente(s)

cancer

Température de stockage

−20°C

Catégories apparentées

Description générale

Changes in the ADP/ATP ratio have been used to differentiate modes of cell death and viability. Increased levels of ATP and decreased levels of ADP signify proliferating cells. Conversely, decreased levels of ATP and increased levels of ADP represent apoptotic or necrotic cells where the decrease in ATP and increase in ADP are much more pronounced in necrosis versus apoptosis.

Application


  • LOC554202 contributes to chordoma progression by sponging miR-377-3p and up-regulating SMAD3.: This article investigates the molecular mechanisms by which LOC554202 facilitates chordoma progression through miR-377-3p and SMAD3 regulation. The ADP/ATP Ratio Assay Kit was utilized to assess the metabolic impact of these molecular changes on cell viability and proliferation (Xu et al., 2023).

  • FOXG1 improves mitochondrial function and promotes the progression of nasopharyngeal carcinoma.: This study explores how FOXG1 enhances mitochondrial function, thereby promoting nasopharyngeal carcinoma progression. The ADP/ATP Ratio Assay Kit was employed to measure mitochondrial function and energy metabolism changes, providing insights into the bioenergetic profile of cancer cells (Xi et al., 2021).

Caractéristiques et avantages

Compatible with high-throughput handling systems.

Adéquation

Suitable for the detection of apoptosis and necrosis in cells and for the studying the effects of compounds on cellular proliferation.

Principe

The ADP/ATP Ratio Assay kit provides a simple and direct procedure for measuring ADP and ATP levels in cells for the screening of apoptosis, necrosis, and cell proliferation. The assay involves two steps. In the first step, the working reagent lyses cells to release ATP and ADP. In the presence of luciferase, ATP immediately reacts with the Substrate D-luciferin to produce light. The light intensity is a direct measure of the intracellular ATP concentration.

Luciferase
ATP + D-Luciferin + O2 ----------> oxyluciferin + AMP + PPi + CO2 + light

In the second step, the ADP is converted to ATP through an enzyme reaction. This newly formed ATP then reacts with the D-luciferin as in the first step. The second light intensity measured represents the total ADP and ATP concentration in the sample.

Produit(s) apparenté(s)

Réf. du produit
Description
Tarif

06693

Timestrip Plus™ -20 °C

Pictogrammes

Exclamation mark
GHS07

Mention d'avertissement

Warning

Mentions de danger

H317,H412

Classification des risques

Aquatic Chronic 3 - Skin Sens. 1

Code de la classe de stockage

10 - Combustible liquids

Faites votre choix parmi les versions les plus récentes :

Certificats d'analyse (COA)

Lot/Batch Number
135CF01A20
135CF01A20
135CE11A26
135CE06A24
135CE06A24

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Retrouvez la documentation relative aux produits que vous avez récemment achetés dans la Bibliothèque de documents.

Consulter la Bibliothèque de documents

Michael St Paul et al.
Cancer immunology research, 8(3), 321-333 (2020-01-23)
CD8+ T cells can be polarized into several different subsets as defined by the cytokines they produce and the transcription factors that govern their differentiation. Here, we identified the polarizing conditions to induce an IL22-producing CD8+ Tc22 subset, which is
Sangiliyandi Gurunathan et al.
Nanomaterials (Basel, Switzerland), 8(6) (2018-06-06)
The unique properties of gold nanoparticles (AuNPs) have attracted much interest for a range of applications, including biomedical applications in the cosmetic industry. The current study assessed the anti-oxidative effect of AuNPs against retinoic acid (RA)-induced loss of cell viability;
Aline G Cozer et al.
Lipids, 51(11), 1303-1307 (2016-10-25)
The present work assesses in vitro the role of human Stanniocalcin 1 (hSTC-1) in glucose metabolism in white retroperitoneal adipose tissue (WRAT) from fed rat. In the fed state, hSTC1 increases the incorporation of
Nidal Zeineh et al.
Cells, 8(7) (2019-07-13)
The 18 kDa translocator protein (TSPO) is an initiator of the mitochondrial apoptosis cascade. Cigarette smoke (CS) exposure provokes alterations in TSPO expression as well as upregulation of its related functions such as mitochondrial membrane potential (ΔψM) and reactive oxygen
Claudia Lin-Kar Hung et al.
Molecular biology of the cell, 29(23), 2809-2820 (2018-09-27)
The huntingtin protein participates in several cellular processes that are disrupted when the polyglutamine tract is expanded beyond a threshold of 37 CAG DNA repeats in Huntington's disease (HD). Cellular biology approaches to understand these functional disruptions in HD have
Afficher tous les articles scientifiques apparentés

Questions

1–6 of 6 Questions  

  1. 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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  2. 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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  3. What is the appropriate concentration range for the assay? Can I use it to test the ADP/ATP ratio in buffer (~ 0.5 - 5 mM ATP and ADP)?

    1 answer

    1. The recommended sample volume is 10 microliters. The higher concentration of 5 mM that was mentioned corresponds to 50 nanomoles. This would be near the upper limit of the assay.
      Bioluminescent ATP assays are very sensitive and accurate over a broad range of the amount of ATP; nanomole and even picomole amounts of ATP can be determined. So, a 5 millimolar sample could be diluted 1000X to 5 micromolar and still give good results, and likely better results than the 5 mM sample.

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  4. Can I please see the product insert and instruction for use for product codeMAK135 ADP/ATP Ratio Assay kit

    1 answer

    1. Here is a link to the product protocol. It can also be found in the 'Documentation' section of the product page: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/135/374/mak135bul.pdf

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  5. Can the cells be stored in a freezer for future use with the MAK135-1KT ADP/ATP Ratio Assay Kit? Should the same approach recommended for tissue testing, involving snap freezing the tissue in liquid nitrogen and deproteinizing it to inactivate residual ATPases, be followed for testing cells?

    1 answer

    1. The approach for cells is similar to that for tissue samples, but with some differences. It is suggested to liquid nitrogen snap freeze the cells in DMSO to prevent cell lysis. Since these are cells, a deproteination step is not necessary. After thawing, the cells should be washed to remove the DMSO and replaced with PBS. Adding a phosphatase inhibitor such as sodium orthovanadate to the PBS can help limit undesired ATPase activity. It is advisable to resuspend the cells in an amount of PBS that allows them to dispense 10^3-10^4 cells in 10 uL sample volumes, as per the ELDT protocol. Quick work is advised, especially after thawing, to initiate the assay, as snap freezing and thawing the cells can stress them. It's preferable to assay freshly harvested cells over snap freezing, as the kit has only been tested on fresh cells.

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  6. How should this item be used on tissue samples? What is the preparation process and what type of buffer should be used?

    1 answer

    1. Given the relatively unstable nature of ATP, if immediate processing of the tissue and running the assay is not feasible after harvesting the tissue, it is advisable to snap freeze the tissue in liquid nitrogen. Prior to utilizing the tissue in the assay, it is recommended to deproteinate the tissue to deactivate any residual ATPases. This process involves initially homogenizing the tissue on ice in PBS + 1 mM EDTA + 0.5% Triton X100, deproteinating samples with TCA, then neutralizing to pH 7 with KOH. Following this, the sample should be centrifuged and the clear supernatant utilized for the assay. An alternative method involves using a 10 kDa spin column for the lysate.

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