Log In Sign up
The largest database of trusted experimental protocols

Ros assay kit

Manufactured by Genmed Scientifics
Visit Supplier
Sourced in China, United States

The ROS assay kit is a laboratory tool used to measure the levels of reactive oxygen species (ROS) in biological samples. It provides a quantitative assessment of ROS, which are important signaling molecules and indicators of oxidative stress in cells.

Automatically generated - may contain errors

Lab products found in correlation

Fluorescence microscope, by Olympus (2 mentions) Dcfh da, by Genmed Scientifics (2 mentions) Fluorescence microplate reader, by Hitachi (1 mentions) Ros assay kit, by Nanjing Jiancheng (1 mentions) Mitosox red, by Thermo Fisher Scientific (1 mentions) Facscalibur, by BD (1 mentions) Firefly luciferase based atp assay kit, by Beyotime (1 mentions) Jc 1 mmp detection kit, by Beyotime (1 mentions) Lipofectamine 3000 reagent, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Sod assay kit, by Keygen Biotech (1 mentions) Gms50009, by Genmed Scientifics (1 mentions) Gms50010, by Genmed Scientifics (1 mentions) Spectramax id5, by Molecular Devices (1 mentions) Atp bioluminescence assay kit, by Beyotime (1 mentions)

10 protocols using ros assay kit

1

AGEs Regulate Schwann Cell ROS

Check if the same lab product or an alternative is used in the 5 most similar protocols
Schwann cells were treated with AGEs only, AGEs + IL-10, or AGEs + NF-κB inhibitor (caffeic acid phenethyl ester, 10 μM, added 30 min before AGE treatment). 3 wells were included in each group.
The cells were cultured for 48 h. The reactive oxygen species (ROS) level was measured using an ROS assay kit (Genmed, Shanghai, China), following the manufacturer's protocol. A plate fluorescence reader (Molecular Devices Corp, Sunnyvale, CA, USA) was used for quantification.
Xu S., Bao W., Men X., Liu Y., Sun J., Li J., Liu H., Cai H., Zhang W., Lou J, & Peng L. (2020). Interleukin-10 Protects Schwann Cells against Advanced Glycation End Products-Induced Apoptosis via NF-κB Suppression. Experimental and clinical endocrinology & diabetes : official journal, German Society of Endocrinology [and] German Diabetes Association, 128(2).
+ Open protocol
+ Expand
2

Kidney Oxidative Stress Measurement

Check if the same lab product or an alternative is used in the 5 most similar protocols
The mice were anesthetized and sacrificed on day 25, and the left kidney was removed and then homogenized as mentioned anteriorly [34 (link)]. The SOD activity and MDA level in plasma and kidney homogenate were detected according to the protocols of corresponding assay kit from Nanjing Jiancheng Bioengineering Institute (Nanjing, China), respectively. The ROS level in kidney homogenate was determined by a commercial ROS assay kit following manufacturer’s protocols (GENMED Scientifics, USA).
Tan L., Tu Y., Wang K., Han B., Peng H, & He C. (2020). Exploring protective effect of Glycine tabacina aqueous extract against nephrotic syndrome by network pharmacology and experimental verification. Chinese Medicine, 15, 79.
+ Open protocol
+ Expand
3

Quantification of Intracellular ROS in Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols
The intracellular ROS of primary cortical neurons was detected using a ROS assay kit (Genmed, Shanghai, China). Briefly, the neurons at DIV 11–13 were incubated with the staining solution (mixture of reagent B and reagent C) for 30 minutes in the dark and washed with Reagent D. Then the fluorescence was detected using a fluorescence microscope (Olympus). The intracellular ROS of N2a neuroblastoma cells was detected using a ROS assay kit (Jiancheng Bioengineering, Nanjing, China). N2a neuroblastoma cells were incubated with 10 μM 2,7-dichlorodi-hydrofluorescein diacetate at 37°C for 30 minutes in the dark, and subjected to the measurement of 2,7-dichlorodi-hydrofluorescein diacetate fluorescence by a fluorescence microplate reader (Hitachi, Tokyo, Japan).
Tao W.Y., Yu L.J., Jiang S., Cao X., Chen J., Bao X.Y., Li F., Xu Y, & Zhu X.L. (2020). Neuroprotective effects of ZL006 in Aβ1–42-treated neuronal cells. Neural Regeneration Research, 15(12), 2296-2305.
+ Open protocol
+ Expand
4

Isolation and Assessment of Hippocampal Mitochondrial ROS

Check if the same lab product or an alternative is used in the 5 most similar protocols
The intact mitochondria of hippocampal region were isolated from brain using a commercial kit (Beyotime, China). The following experimental procedures were conducted according to the manufacturer’s instructions. Briefly, the homogenate was centrifuged at 6,000 g at 4°C for 5 min. The collected supernatant was further centrifuged at 11,000 g at 4°C for 10 min to obtain a mitochondrial pellet. Then, the mitochondrial ROS was detected utilizing the ROS assay kit (Genmed Scientifics, Shanghai, China).
Cellular mitochondrial ROS activity was assessed with Mito SOX Red (Invitrogen) staining. BV2 cells were seeded onto six-well blank plates with a density of 1 × 105/ml with 3 parallel wells in each group. Cells were incubated with MitoSOX Red probe at a final concentration of 5 μM for 10 min at 37°C and washed twice with PBS. Quantification of mtROS release was conducted by FACSCalibur (BD Biosciences). All data were analyzed on FlowJo software (Tree Star, San Carlos, CA).
Wu X., Gong L., Xie L., Gu W., Wang X., Liu Z, & Li S. (2021). NLRP3 Deficiency Protects Against Intermittent Hypoxia-Induced Neuroinflammation and Mitochondrial ROS by Promoting the PINK1-Parkin Pathway of Mitophagy in a Murine Model of Sleep Apnea. Frontiers in Immunology, 12, 628168.
+ Open protocol
+ Expand
5

Measurement of Cellular Bioenergetics

Check if the same lab product or an alternative is used in the 5 most similar protocols
ATP content was measured with a firefly luciferase-based ATP assay kit (Beyotime Institute of Biotechnology, Shanghai, China). MMP levels were determined with a 5,5′,6,6′-tetrachloro-1,1′,3,3′ tetraethylbenzimidazolylcarbocyanine iodide (JC-1) MMP detection kit (Beyotime Institute of Biotechnology, Shanghai, China), and intracellular ROS were detected with a ROS assay kit (Genmed Scientifics Inc., Shanghai, China) containing an oxidation-sensitive fluorescent probe (DCFH-DA). All measurements were performed according to the manufacturer’s instructions. Data were normalized to the control group values and expressed as a percentage of control levels.
Zuo Y., Yin L., Cheng X., Li J., Wu H., Liu X., Gu E, & Wu J. (2020). Elamipretide Attenuates Pyroptosis and Perioperative Neurocognitive Disorders in Aged Mice. Frontiers in Cellular Neuroscience, 14, 251.
+ Open protocol
+ Expand
6

Quantifying Oxidative Stress in HepG2 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
The HepG2 hepatocellular line was purchased from Classic Specimen Culture and Storage Center at Wuhan University (Wuhan, China). The cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 1% penicillin/streptomycin and 10% FBS (Gibco, Thermo Fisher, Waltham, MA, USA) in 5% CO 2 at 37°C in a humidified atmosphere. Cells (5 × 10 5 cells per well in 6-well culture plates) were transfected with the pWPI-MsrA-GFP vector or the control pWPI-GFP vector using Lipofectamine 3000 reagent (Invitrogen, Waltham, MA, USA) according to the manufacturer's instructions. After transfected with vectors for 48 hours, intracellular ROS levels were detected by the fluorescent dye dihydroethidium bromide (DHE) using a ROS assay kit (GENMED Scientifics Inc, Arlington, MA, USA) according to the manufacturer's protocols. GFP and red DHE fluorescence were imaged with a fluorescence microscope (Olympus, Tokyo, Japan). The intracellular ROS levels were measured using Image-Pro Plus 6.0 software, represented by the mean DHE fluorescence intensity in the detected area of cells, and normalized to untreated control. Meanwhile, the mRNA levels of interleukin-6 (IL-6) and tumor necrosis factor α (TNFα) were determined by real-time quantitative RT-PCR (qRT-PCR).
Xu Y., Li F., Zhao X., Tan C., Wang B., Chen Y., Cao J., Wu D, & Yu H. (2020). Methionine sulfoxide reductase A attenuates atherosclerosis via repairing dysfunctional HDL in scavenger receptor class B type I deficient mice. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 34(3).
+ Open protocol
+ Expand
7

Hippocampal Oxidative Stress in Postnatal Rats

Check if the same lab product or an alternative is used in the 5 most similar protocols
Rats (n = 6 for each group) were sacrificed immediately postanesthesia (at PND 7), and the hippocampi were removed quickly. Intracellular ROS were detected using a ROS assay kit (Genmed Scientifics Inc., Shanghai, China) containing an oxidation-sensitive fluorescent probe (DCFH-DA) with a spectrofluorometer (excitation 490 nm, emission 520 nm). Malondialdehyde (MDA) is an end-product of ROS-induced peroxidation. Superoxide Dismutase (SOD) is an important enzyme that participates in the removal of ROS from the cellular environment. The extent of lipid peroxidation was estimated by MDA levels, which were measured by using the spectrophotometric diagnostic kits (Jiancheng Biological Technology Co., Ltd., Nanjing, China) according to the manufacturer’s instructions. The SOD activity was determined using a SOD assay kit (Jiangsu KeyGEN BioTECH Co., Ltd., Nanjing, China) according to manufacturer’s instructions. Enzyme activity was converted to units per milligram of protein. One unit of SOD activity was defined as the amount that reduced the absorbance at 550 nm by 50%.
Wu J., Hao S., Sun X.R., Zhang H., Li H., Zhao H., Ji M.H., Yang J.J, & Li K. (2017). Elamipretide (SS-31) Ameliorates Isoflurane-Induced Long-Term Impairments of Mitochondrial Morphogenesis and Cognition in Developing Rats. Frontiers in Cellular Neuroscience, 11, 119.
+ Open protocol
+ Expand
8

Mitochondrial ROS and Antioxidant Enzyme Activity

Check if the same lab product or an alternative is used in the 5 most similar protocols
The mtROS were measured using the ROS Assay Kit (GMS 10104.1, GenMed Scientifics, Shanghai, China) according to the manufacturer's instructions. The serum and hippocampus SOD activity were measured using the SOD Assay Kit (Jiacheng Biological Technology Co., Ltd., Nanjing, China) according to the manufacturer's instructions. The enzyme activity was converted to units per milligram of protein. One unit of SOD activity was defined as the amount of that reduced the absorbance at 450 nm by 50%. The levels of electron transport chain (ETC) components—I/III/IV were also measured using the ETC Analyzing Kit (GMS 50007 for ETC-I, GMS 50009 for ETC-III and GMS 50010 for ETC-IV, GenMed Scientifics, Shanghai, China). All the above procedures were conducted on ice in a dark room. The results were collected by a microplate reader (SpectraMax iD5, Molecular Devices, San Jose, CA, USA).
Liu Y., Fu H., Wu Y., Nie B., Liu F., Wang T., Xiao W., Yang S., Kan M, & Fan L. (2021). Elamipretide (SS-31) Improves Functional Connectivity in Hippocampus and Other Related Regions Following Prolonged Neuroinflammation Induced by Lipopolysaccharide in Aged Rats. Frontiers in Aging Neuroscience, 13, 600484.
+ Open protocol
+ Expand
9

ROS and ATP Quantification Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
Intracellular ROS was detected using a ROS assay kit (Genmed Scientifics Inc.) containing an oxidation-sensitive fluorescent probe (DCFH-DA) in a spectrofluorometer (excitation 490 nm, emission 520 nm). Assessment of relative ATP contents was performed using the ATP bioluminescence assay kit (Beyotime Institute of Biotechnology) following the manufacturer’s instructions.
Wu J., Li H., Sun X., Zhang H., Hao S., Ji M., Yang J, & Li K. (2015). A Mitochondrion-Targeted Antioxidant Ameliorates Isoflurane-Induced Cognitive Deficits in Aging Mice. PLoS ONE, 10(9), e0138256.
+ Open protocol
+ Expand
10

Oxidative Stress Evaluation after ROSC

Check if the same lab product or an alternative is used in the 5 most similar protocols
To detect the oxidative stress of the brain tissue at 24 hours after ROSC, immunohistochemical staining of 4-hydroxynonenal (4-HNE) and 8-hydroxy-2′-deoxyguanosine (8-OHdG) was conducted. 4-HNE was conducted to assess lipid peroxidation while 8-OHdG was conducted to evaluate the extent of nucleic acid oxidation. For immunohistochemical staining, the brain sections were incubated with a primary monoclonal antibody against 4-HNE and 8-OHdG (Abcam, Cambridge, United Kingdom) overnight at 4°C. After primary antibody incubation, sections were washed three times for 10 minutes with phosphate-buffered saline. After washing, they were incubated with the corresponding secondary antibody for 2 hours at room temperature. Six fields of vision (400x magnification) of these sections were selected randomly and photographed with an optical microscope. For analysis, the AOD of 4-HNE and 8-OHdG were calculated using Image J software (National Institute of Health, United States). The ROS levels in the brain at 6 hours after ROSC were measured using a ROS assay kit (Genmed Scientifics, Wilmington, USA) as the manufacturer's instructions.
Tan Y., Zhang J., Ge Q., Fang X., Song F., Yu T., Jiang L., Wei Y, & Wang P. (2022). Ketone Body Improves Neurological Outcomes after Cardiac Arrest by Inhibiting Mitochondrial Fission in Rats. Oxidative Medicine and Cellular Longevity, 2022, 7736416.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!