Tissue samples must be prepared as fresh or frozen extracts. PFA-embedded tissues are not suitable. Yes , PFA may interfere with the Glutathione Assay. Please see the link below to review additional information available in the product datasheet:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/146/686/mak440pis-ms.pdf
Przejdź do
MAK440
Glutathione GSH/GSSG Assay Kit
Sufficient for 100 colorimetric tests
Synonim(y):
GSH/GSSG Ratio Kit
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| Do Państwa/SKU | Dostępność | Cena netto |
|---|---|---|
1 kit | Skontaktuj się z Obsługą Klienta, aby uzyskać informacje na temat dostępności | 1860,00 zł |
Informacje o tej pozycji
NACRES:
NA.84
UNSPSC Code:
12161503
1860,00 zł
Skontaktuj się z Obsługą Klienta, aby uzyskać informacje na temat dostępności
Pomoc techniczna
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Pozwól nam pomócQuality Segment
application(s)
pharmaceutical
detection method
colorimetric
relevant disease(s)
Alzheimer′s disease; Parkinson′s disease; cancer; neurological disorders; aging/geriatric diseases
storage temp.
−20°C
General description
Glutathione (GSH), composed of the amino acids glycine, glutamic acid, and cysteine, serves as a fundamental antioxidant within the body, playing a pivotal role in cellular defense against oxidative stress caused by reactive oxygen species (ROS). As the major free thiol in cells, GSH is instrumental in various biological processes, including the detoxification of harmful substances, the removal of hydroperoxides, and the maintenance of protein sulfhydryl groups in their reduced state. GSH achieves this by directly reducing disulfide bonds in cytoplasmic proteins to cysteines, during which it is transformed into its oxidized form, glutathione disulfide (GSSG). This cycle between reduced and oxidized forms is critical for maintaining the cellular redox balance.Glutathione functions as a cofactor for several key antioxidant enzymes, enhancing their ability to neutralize free radicals. It also plays a crucial role in the regeneration of vitamins C and E, thereby contributing to their antioxidative effects. Beyond its antioxidant properties, GSH is vital for mitochondrial function and the integrity of mitochondrial DNA (mtDNA), which is essential for energy production and cellular health. Dysregulation of glutathione levels is associated with the pathogenesis of several neurodegenerative diseases such as Alzheimer′s, Parkinson′s, and Huntington′s diseases, as well as other health conditions including cystic fibrosis, immune disorders, and cardiovascular diseases. Glutathione′s role in metabolism and cellular protection is extensive, influencing cellular proliferation, apoptosis, and overall physiological stability.
Application
The Glutathione GSH/GSSG Assay Kit may be used for:
- Cancer Research
- Diabetes Research
- Neurodegenerative Disease Research
The kit is suitable for the quantitative determination of reduced and oxidized glutathione (GSH/GSSG) in whole blood, plasma, serum, urine, tissue and cell extracts.
Biochem/physiol Actions
The Glutathione GSH/GSSG Assay Kit is designed to accurately measure total, reduced and oxidized glutathione in biological samples using an enzymatic method that utilizes Ellman′s Reagent (DTNB) and glutathione reductase (GR). DTNB reacts with reduced glutathione to form a yellow product. The rate of change in the optical density, measured at A412 nm, is directly proportional to the glutathione concentration in the sample. This kit can also be used to measure oxidized (GSSG) by using a specific protocol which first scavenges all GSH with 1-methyl-2-vinylpyridinium triflate.
Features and Benefits
Sensitive Detection Range: Detect Glutathione levels accurately across a wide range, from 0.01 µM to 3 µM, using only 25 μL of sample in a 96-well plate setup, ensuring highly sensitive analysis with minimal sample requirement.
Simplified Process: Experience a streamlined process with the addition of only a single working reagent and a 10 minute room temperature reaction, reducing complexity and saving valuable time and effort.
Compatibility with High-Throughput Systems: Easily incorporate our kit into high-throughput handling systems, ensuring smooth and accurate processing, enhancing efficiency in your laboratory workflow.
Simplified Process: Experience a streamlined process with the addition of only a single working reagent and a 10 minute room temperature reaction, reducing complexity and saving valuable time and effort.
Compatibility with High-Throughput Systems: Easily incorporate our kit into high-throughput handling systems, ensuring smooth and accurate processing, enhancing efficiency in your laboratory workflow.
Other Notes
For additional information on our range of Biochemicals, please complete this form.
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Ta pozycja | |||
|---|---|---|---|
| description Sufficient for 100 colorimetric tests | description Sufficient for 100 colorimetric tests | description Sufficient for 100 colorimetric tests | description sufficient for 700 assays |
| Quality Level 100 | Quality Level 200 | Quality Level 200 | Quality Level 200 |
| detection method colorimetric | detection method colorimetric | detection method colorimetric | detection method - |
| storage temp. −20°C | storage temp. 2-8°C | storage temp. −20°C | storage temp. 2-8°C |
| application(s) pharmaceutical | application(s) - | application(s) - | application(s) - |
| relevant disease(s) Alzheimer′s disease; Parkinson′s disease; cancer; neurological disorders; aging/geriatric diseases | relevant disease(s) - | relevant disease(s) - | relevant disease(s) - |
Klasa składowania
10 - Combustible liquids
flash_point_f
188.6 °F
flash_point_c
87 °C
wgk
WGK 2
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Can tissue samples be used embedded in PFA or do they need to be frozen down/ fresh for the assay? Can PFA interfere in any way?
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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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How should I prepare a tissue sample for this assay? How much tissue sample to start with? Should I just take tissue, add with the cold buffer as mentioned for cell lysate and sonicate?
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Here is the protocol to prepare tissue samples for this kit: Start with 20-100 mg tissue, add 200-1000 μL ice-cold PBS. Lysis can be achieved by homogenization (10-20 passes in a Dounce homogenizer on ice) or by sonication (preferably performed in an ice-water bath). The degree of tissue lysis can be checked under a microscope. Centrifuge homogenate at 14,000 g for 10 min. Transfer the clear supernatant into a clean tube. It is prudent to run a pilot test of the sample at different dilutions. Choose a dilution with the readings in the detection range of the standard curve for further assays.
Please note that β-mercaptoethanol, dithiothreitol and cysteine are known to interfere in this assay so it is important to avoid the use of these compounds during sample preparation.
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How to make 5% MPA? The MPA recommended in the manual (239275) is 35% pure. Does 5%(w/v) refer to the concentration of pure MPA or the impure solid mixture? In this case do I need to account for the 35%?
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The 5% MPA solution should be made without considering purity (i.e. 5 g of cat#239275 in 100 g of water). Please note that the 5% MPA should be prepared fresh each day it is to be used.
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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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Hi, Can I use this kit for testing GSH/GSSG ratio in plants (Arabidopsis), if yes, is there any modifications in the protocol for plants? Thank you
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This kit can work on whole blood, plasma, serum, urine, tissue, and cell extracts samples; however, it has not been tested for plant samples, including Arabidopsis. Therefore, no specific modification in the protocol is made to test the GSH/GSSG ratio in plants using this kit.
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After the extraction procedure on colon tissues, do I have to also realize the deproteination before the assay ?
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Colon tissue samples must undergo the deproteination procedure as described on page 2 of the kit's technical bulletin.
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Can this kit be used to analyze plasma or serum samples? If yes, is plasma or serum better? If yes, what is the procedure to follow as the procedure is described for whole blood and cell lysate only. What is the stability of frozen plasma for glutathione?
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Serum or plasma samples can be prepared using the same method as whole blood samples. Both types of samples are suitable for testing and are equally recommended. EDTA plasma is specifically known to be compatible with the assay.
Samples of serum or plasma can be frozen for later analysis; however, it is advisable to freeze them promptly after preparation at -80°C. To minimize the risk of glutathione degradation, avoid multiple freeze-thaw cycles and aim to test the samples within one month.
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Where does the DF of the calculation come from when using Scavenger or without Scavenger? If I need to make an additional dilution do I multiply it?
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The volumes used for buffer and scavenger should not be altered from the stated volumes.
For all samples treated with Scavenger, DF = 165. For samples not treated with Scavenger, DF = 450 for whole blood and 150 for all other samples. Please refer to the technical bulletin, specifically page 4, for information on the calculation of GSH and GSSG.Helpful?
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How is the test for GSSG and total GSH performed when using dry biomass? Is only PBS used or should phosphate buffer + EDTA be added? What is the estimated sonication time?
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In this case, the use of PBS is not recommended. For preparation of a sample from the dry biomass, follow the procedures as described for preparing a cell lysate. That is, use 50 mM phosphate, pH 7, 1 mM EDTA, and 20 microliters of Scavenger for preparation of the sample for GSSG measurement, and use 50 mM phosphate, pH 7, and 1 mM EDTA (with NO scavenger) for preparation of the sample for GSH measurement. Sonication for 30-60 seconds should be sufficient, but examination of a sample under a microscope to ensure that it is completely homogenized is recommended.
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