Intracellular Total ROS Activity Assay Kit
Intracellular Total ROS Activity Assay Kit
Applications
Flow Cytometer
Synonyms
Intracellular Total ROS Activity; Intracellular Total ROS Activity Assay Kit; Intracellular Total ROS Activity Assay; Intracellular Total ROS Activity assay kit
Applicable Applications for Intracellular Total ROS Activity assay kit
Flow Cytometer
Application Notes
1. Prepare samples and controls in 1X Assay Buffer or buffer of choice (0.5 mL at a cell density of 5 x 105-1 x 106 cells/mL).
2. Reconstitute Total ROS Green with 100 uL DMSO to produce a 500X stock concentrate.
3. Dilute 1:10 by adding 900 uL 1X Assay Buffer to the 500X stock Total ROS Green to produce a 50X working solution.
4. Add Total ROS Green to each sample at 1:50. For example, add 10 uL into 490 uL cultured cells.
5. Incubate 1 hour at 37 degree C.
6. Treat cells with test compounds for desired period of time. Keep cells protected from light.
7. Analyze with a flow cytometer. Total ROS Green excites at 490 nm and emits at 520 nm.
2. Reconstitute Total ROS Green with 100 uL DMSO to produce a 500X stock concentrate.
3. Dilute 1:10 by adding 900 uL 1X Assay Buffer to the 500X stock Total ROS Green to produce a 50X working solution.
4. Add Total ROS Green to each sample at 1:50. For example, add 10 uL into 490 uL cultured cells.
5. Incubate 1 hour at 37 degree C.
6. Treat cells with test compounds for desired period of time. Keep cells protected from light.
7. Analyze with a flow cytometer. Total ROS Green excites at 490 nm and emits at 520 nm.
Sample
Cell culture
Excitation/Emission
490 nm / 520 nm
Preparation and Storage
Store total ROS Green and DMSO at < = -20 degree C. Store Assay Buffer at < = 2-8 degree C.
Related Product Information for Intracellular Total ROS Activity assay kit
Description: Intracellular Total ROS Activity Assay provides a good screening option for assessing the potency of oxidative stress inhibitor and activator reagents, and will help to determine how oxidative stress modulates varied intracellular pathways. This product assesses the overall level of intracellular ROS activity and can be analyzed using a flow cytometer.
Background: Reactive oxygen species (ROS) are natural byproducts of the normal metabolism of oxygen and play important roles in cell signaling. However, during oxidative stress-related states, ROS levels can increase dramatically. The accumulation of ROS results in significant damage to cell structures. The role of oxidative stress in cardiovascular disease, diabetes, osteoporosis, stroke, inflammatory diseases, a number of neurodegenerative diseases and cancer, has been well established.
Intracellular Total ROS Activity Assay provides a good screening option for assessing the potency of oxidative stress inhibitor and activator reagents, and will help to determine how oxidative stress modulates varied intracellular pathways. This kit assesses the overall level of intracellular ROS activity, but does not identify the specific reactive oxygen molecule(s) generated by the oxidative stress event.
The key reagent in the assay is a proprietary dye called Total ROS Green. This dye quickly penetrates membrane structures and accumulates within the cell. In the presence of ROS, the non-fluorescent Total ROS Green dye molecule is oxidized by all various iterations of ROS molecular forms. In the oxidized state, the Total ROS Green dye molecule acquires fluorescence properties that enable its detection by flow cytometry as an indicator of the relative level of intracellular ROS activity.
Background: Reactive oxygen species (ROS) are natural byproducts of the normal metabolism of oxygen and play important roles in cell signaling. However, during oxidative stress-related states, ROS levels can increase dramatically. The accumulation of ROS results in significant damage to cell structures. The role of oxidative stress in cardiovascular disease, diabetes, osteoporosis, stroke, inflammatory diseases, a number of neurodegenerative diseases and cancer, has been well established.
Intracellular Total ROS Activity Assay provides a good screening option for assessing the potency of oxidative stress inhibitor and activator reagents, and will help to determine how oxidative stress modulates varied intracellular pathways. This kit assesses the overall level of intracellular ROS activity, but does not identify the specific reactive oxygen molecule(s) generated by the oxidative stress event.
The key reagent in the assay is a proprietary dye called Total ROS Green. This dye quickly penetrates membrane structures and accumulates within the cell. In the presence of ROS, the non-fluorescent Total ROS Green dye molecule is oxidized by all various iterations of ROS molecular forms. In the oxidized state, the Total ROS Green dye molecule acquires fluorescence properties that enable its detection by flow cytometry as an indicator of the relative level of intracellular ROS activity.
Product Categories/Family for Intracellular Total ROS Activity assay kit
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Product Notes
The Intracellular Total ROS Activity (Catalog #AAA258376) is an Assay Kit and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's Intracellular Total ROS Activity can be used in a range of immunoassay formats including, but not limited to, Flow Cytometer. 1. Prepare samples and controls in 1X Assay Buffer or buffer of choice (0.5 mL at a cell density of 5 x 105-1 x 106 cells/mL). 2. Reconstitute Total ROS Green with 100 uL DMSO to produce a 500X stock concentrate. 3. Dilute 1:10 by adding 900 uL 1X Assay Buffer to the 500X stock Total ROS Green to produce a 50X working solution. 4. Add Total ROS Green to each sample at 1:50. For example, add 10 uL into 490 uL cultured cells. 5. Incubate 1 hour at 37 degree C. 6. Treat cells with test compounds for desired period of time. Keep cells protected from light. 7. Analyze with a flow cytometer. Total ROS Green excites at 490 nm and emits at 520 nm. Researchers should empirically determine the suitability of the Intracellular Total ROS Activity for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Intracellular Total ROS Activity, Assay Kit" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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