Oxidative Stress Assay
Sensitive, Reproducible & Easy-to-use Oxidative stress assays
The follwing assays are available to measure cellular Cytotoxicity:
ROS Detection Assay
Advantages & Features
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Based on unique fluorescent probe H2DCFDA.
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Fast & Easy Protocol.
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Safe: non-Radioactive, no special handling needed.
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High-Throughput.
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Accurate & Reproducible results.
Overview:
ROS Detection Assay Kit is a highly sensitive and safe assay to measure Reactive Oxygen Species. ROS include a number of reactive molecules and free radicals, derived from molecular oxygen, that damage DNA and RNA and oxidize proteins and lipids (lipid peroxidation). The most common ROS include superoxide anion (O2•-), hydrogen peroxide (H2O2), hydroxyl radical (HO•), and singlet oxygen (1O2), all of which are more reactive than oxygen (O2) itself. The molecules are produced during the electron transport of mitochondrial aerobic respiration or by oxidoreductase enzymes and metal-catalyzed oxidation.
It uses the cell-permeant reagent Dichlorodihydrofluorescein-diacetate (H2DCFDA), a fluorogenic dye that measures hydroxyl, peroxyl, and other reactive oxygen species (ROS) activity within the cell. After diffusion into the cell, the acetyl groups on H2DCFDA are cleaved by intracellular esterase to yield the non-fluorescent compound which is rapidly oxidized to highly fluorescent 2’,7’-Dichlorodihydrofluorescein by ROS. The fluorescence intensity is proportional to the ROS levels within the cell cytosol.
Applications
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Measurement of intracellular levels of ROS.
Content
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5 mg Fluorescent Substrate (H2DCFDA)
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50 mL 10X Assay Buffer
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500 μL Positive control (H2O2, 8.8M)
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1 mL DMSO
Storage
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Shipment Gel Pack.
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Storage: 4 °C upon receipt.
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Shelf Life: 24 months from the date of purchase, if it is properly stored.
TBARS Assay
Advantages & Features
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Accurate.
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Simple & Fast Protocol.
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Versatile: Colorimetric or Fluorometric MDA Quantitation.
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Standardized tool for assaying lipid peroxidation in Plasma, Serum, Urine, Tissue homogenates, and Cell Lysates.
Overview:
Oxidative stress in the cellular environment results in the formation of highly reactive and unstable lipid hydroperoxides. Decomposition of the unstable peroxides derived from polyunsaturated fatty acids results in the formation of malondialdehyde (MDA), which can be quantified colorimetrically following its controlled reaction with thiobarbituric acid. Thiobarbituric Acid Reactive Substances (TBARS) assay was proposed over 40 years ago and is now the most commonly used of method to screening and monitoring lipid oxidation.
TBARS method has been used to evaluate a wide range of samples that include human and animal tissues and fluids, drugs, foods and natural products. The sensitivity of measuring Thiobarbituric Acid Reactive Substances (TBARS) has made this assay the method of choice for screening and monitoring lipid peroxidation, a major indicator of oxidative stress. Even though there remains a controversy cited in literature regarding the specificity of TBARS toward compounds other than MDA, it still remains the most widely employed assay used to determine lipid peroxidation.
This assay is based on the reaction of malondialdehyde (MDA) with thiobarbituric acid (TBA); TBA reacts with MDA to form a pink chromogen, which can be detected spectrophotometrically at 532 nm.
Applications
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For quantitative determination of lipid Peroxides (thiobarbituric acid reactive substances, TBARS).
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Evaluation of drug effects on lipid Peroxidation.
Content
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– 4 x 0.53 g Thiobarbituric Acid (TBA)
– 40 mL Diluent 1 (contain Acetic Acid)
– 20 mL Diluent 2 (contain NaOH)
– 10 mL SDS Solution
– 250 μL MDA Standard (10mM)
– 20 mL H2O2 solution
– 1 unit 96-Well Solid Black plate
– 1 unit 96-Well Solid Clear plate
– 2 Adhesive strips
Storage
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Shipment: Gel Pack.
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Storage: 4 °C upon receipt.
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Shelf Life: 24 months from the date of purchase, if it is properly stored.