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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.pdf
Przejdź do
MAK135
ADP/ATP Ratio Assay Kit
sufficient for 100 tests (bioluminescent)
Wybierz wielkość
| Do Państwa/SKU | Dostępność | Cena netto |
|---|---|---|
1 kit | Skontaktuj się z Obsługą Klienta, aby uzyskać informacje na temat dostępności | 1920,00 zł |
Informacje o tej pozycji
1920,00 zł
usage
sufficient for 100 tests (bioluminescent)
application(s)
pharmaceutical
detection method
chemiluminescent
relevant disease(s)
cancer
storage temp.
−20°C
General description
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).
Biochem/physiol Actions
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.
Features and Benefits
1 of 1
Ta pozycja | |||
|---|---|---|---|
| description sufficient for 100 tests (bioluminescent) | description sufficient for 100 assays (bioluminescent) | description sufficient for 200 colorimetric tests | description Sufficient for 200 tests |
| detection method chemiluminescent | detection method chemiluminescent | detection method colorimetric | detection method - |
| usage sufficient for 100 tests (bioluminescent) | usage sufficient for 100 assays (bioluminescent) | usage sufficient for 200 colorimetric tests | usage - |
| storage temp. −20°C | storage temp. −20°C | storage temp. 2-8°C | storage temp. −20°C |
| application(s) pharmaceutical | application(s) - | application(s) - | application(s) - |
| relevant disease(s) cancer | relevant disease(s) hematological disorder; cancer | relevant disease(s) cancer | relevant disease(s) - |
signalword
Warning
hcodes
Hazard Classifications
Aquatic Chronic 3 - Skin Sens. 1
Klasa składowania
10 - Combustible liquids
wgk
WGK 3
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How can I determine the shelf life / expiration / retest date of this product?
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Once the kit is opened, can it be frozen again for further analysis or must the entire kit be used once opened?
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The cosubstrate is not highly sensitive to freeze-thaw cycles; however, the substrate should not undergo multiple freeze-thaw cycles. Reagents can be aliquoted and frozen after the first thaw without significant issues expected. If enzyme components reach elevated temperatures during assay preparation, the signal may be adversely affected. It is recommended to keep enzyme aliquots on ice until the ATP and ADP reagents are prepared.
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Ce kit peut-il être appliqué aux tissus ?
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This kit was originally designed to measure the ADP/ATP ratio in cultured animal cells. It is not suitable for direct use on fabric samples. But since ADP and ATP are very water soluble, the kit presumably could be used on a sample that is an aqueous extract of a fabric sample.
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Can I use this kit for a yeast that produced an orange-red carotenoid? Could this interfere at the results?
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The effect of carotenoids on assay results has not been tested. It is possible that carotenoids could affect the results of this assay. A non-polar solvent such as hexane or dichloromethane may be used to sequester the carotenoids in the solvent, leaving the ATP and ADP in the aqueous phase. In this case, the clarified aqueous phase should be used as the sample.
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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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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)?
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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.Helpful?
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Can I please see the product insert and instruction for use for product codeMAK135 ADP/ATP Ratio Assay kit
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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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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?
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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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How should this item be used on tissue samples? What is the preparation process and what type of buffer should be used?
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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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