Yes, this kit can be used to detect ROS in bacteria. The kit has been tested on hydrogen peroxide (H2O2), hydroxyl radical (•OH), tert-butyl-hydroperoxide (TBHP), and hypochlorous acid (HOCl). Kit MAK142 has also been tested on superoxide anions (O2•-), but MAK143 does not.
MAK143
Fluorometric Intracellular Ros Kit
sufficient for 200 fluorometric tests (green)
Synonym(s):
Fluorometric ROS Detection Kit
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About This Item
NACRES:
NA.25
UNSPSC Code:
12161503
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Product Name
Fluorometric Intracellular Ros Kit, sufficient for 200 fluorometric tests (green)
usage
sufficient for 200 fluorometric tests (green)
detection method
fluorometric
relevant disease(s)
cancer; neurological disorders
storage temp.
−20°C
Related Categories
1 of 4
This Item | MAK144 | MAK142 | MAK145 |
|---|---|---|---|
| detection method fluorometric | detection method fluorometric | detection method fluorometric | detection method fluorometric |
| usage sufficient for 200 fluorometric tests (green) | usage sufficient for 200 fluorometric tests (orange) | usage sufficient for 200 fluorometric tests (Deep red) | usage sufficient for 200 fluorometric tests (red) |
| storage temp. −20°C | storage temp. −20°C | storage temp. −20°C | storage temp. −20°C |
| relevant disease(s) cancer; neurological disorders | relevant disease(s) cancer; neurological disorders | relevant disease(s) neurological disorders; cancer | relevant disease(s) neurological disorders; cancer |
Application
Fluorometric Intracellular Ros Kit has been used in reactive oxygen species (ROS) generation assay.[1]
This kit is suitable for the detection of intracellular ROS by fluorescence microplate reader or a fluorescent microscope.
Biochem/physiol Actions
The Fluorometric Intracellular ROS Assay Kit provides a sensitive, one-step fluorometric assay to detect intracellular ROS (especially superoxide and hydroxyl radicals) in live cells within 1 hour incubation. ROS react with a fluorogenic sensor localized to the cytoplasm, resulting in a fluorometric product (λex = 490/λem = 520 nm) proportional to the amount of ROS present. The assay can be performed in either a 96 or 384 multiwell plate format and read using a fluorescence microscope or a fluorescence microplate reader.
General description
Reactive oxygen species (ROS) are generated as a result of the reduction of oxygen during aerobic respiration and by various enzymatic systems within the cell. At physiological levels, ROS contribute to cell signaling and host defense. Increased ROS generation, above the detoxification capacity of the biological system, results in oxidative stress and cellular damage. The main damage to cells results from the ROS-induced alteration of macromolecules such as polyunsaturated fatty acids in membrane lipids, essential proteins, and DNA. ROS has been implicated in disease states, such as Alzheimer′s disease, Parkinson′s disease, cancer, and aging.
Storage Class
10 - Combustible liquids
wgk
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
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Instructions
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Does this test detect ROS in Gram-positive bacteria? Which ROS does it detect? Only superoxide or all of them?
1 answer-
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On the technical bulletin it says that i have use 11x test compound. What does it means? The reagent is sensitive to small ROS production? At concentration 11x my cells would immediately die
1 answer-
The test compounds referenced in the technical bulletin for this product are the species being tested for stimulation or inhibition of intracellular reactive oxygen species (ROS) production. The kit has been calibrated so that 11 times the desired working concentration of test compounds should be used with a 96-well format, and 6 times the desired working concentration should be used with a 384-well format.
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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 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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Does the fluorescence produced by ROS resist PFA fixation? Is it detergent resistant?
1 answer-
This kit is intended to be used on living cultured cells. It is not recommended for use on paraformaldehyde (PFA) fixed cells or tissue, or on cells or tissue that have been treated with a detergent or surfactant.
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What is the mechanism for detecting ROS? Specifically, what component in the ROS Detection Reagent enables the detection of ROS upon their formation?
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The component A ROS green is a non-conjugated compound that is concealed with a ROS-cleavable blocking group. Upon the ROS reaction, the blocking group is cleaved by ROS to release a strongly fluorescent dye. The ROS production is proportional to the concentration of the fluorophore.
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Is ROS Detection Reagent(Green) the same as 2′,7′-Dichlorofluorescin diacetate?
1 answer-
The composition information for the ROS Detection Reagent, Green solution is considered prioprietary.
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