Oxidative Stress Assay Kits

Total Antioxidant Capacity Assay Kit
ABL-TAC-Peroxyl-KI01

• Inhibition assay of chemiluminescence caused by peroxyl radicals.

• Includes peroxyl radical generator that mimics natural lipid peroxidation.

• Applicable to serum, semen plasma, CSF, tissue homogenates, urine, tea, fruits, wines, juices, botanical extracts.

This assay is the most popular method for the analysis of a wide range of biological samples such as serum (plasma), CSF, semen plasma, tissue homogenates and urine. It can also been used in the analysis of tea, wine, fruits and botanical extracts. The platform of this kit is an artificial system where biologically relevant peroxyl free radicals are generated by thermal decomposition of 2,2-azobis(2-amidinopropane) (ABAP). The ABAP decomposition products are a pair of C-centered free radicals R and a nitrogen molecule. The R free radicals further react with oxygen molecules to form peroxyl radicals ROO, which are similar to those found in vivo during lipid peroxidation. These peroxyl radicals react with an indicator molecule, luminol (LH2), to generate a luminol radical (LH) that results in emission of blue lights centered at ~425 nm. When antioxidants are present, such a light production is inhibited until the antioxidants are exhausted. The time of inhibition or the induction time to light production is proportional to the total concentration of antioxidants. The antioxidant concentration is determined by comparing induction time to that of a water-soluble vitamin E (tocopherol) analog, Trolox. 

Antioxidant Assay Kit 

CAY-709001-KI01

Catalase Assay Kit

CAY-707002

Catalase (EC 11.6; 2H2O2 oxidoreductase) is an ubiquitous antioxidant enzyme that is present in most aerobic cells. Catalase (CAT) is involved in the detoxification of hydrogen peroxide (H2O2), a reactive oxygen species (ROS), which is a toxic product of both normal aerobic metabolism and pathogenic ROS production. This enzyme catalyzes the conversion of two molecules of H2O2 to molecular oxygen and two molecules of water (catalytic activity). CAT also demonstrates peroxidatic activity, in which low molecular weight alcohols can serve as electron donors. While the aliphatic alcohols serve as specific substrates for CAT, other enzymes with peroxidatic activity do not utilize these substrates. In humans, the highest levels of catalase are found in liver, kidney, and erythrocytes, where it is believed to account for the majority of hydrogen peroxide decomposition.
The Cayman Chemical Catalase Assay Kit utilizes the peroxidatic function of CAT for determination of enzyme activity. The method is based on the reaction of the enzyme with methanol in the presence of an optimal concentration of H2O2. The formaldehyde produced is measured spectrophotometrically with 4-amino-3-hydrazino-5-mercapto-1,2,4-triazole (Purpald) as the chromogen. Purpald specifically forms a bicyclic heterocycle with aldehydes, which upon oxidation changes from colorless to a purple color. The assay can be used to measure CAT activity in plasma, serum, erythrocyte lysates, tissue homogenates, and cell lysates.

Catalase Activity Assay Kit
ABL-CAT-240-KI01 1 Kit

• Colorimetric Assay Kit.

• Based on hydrogen peroxide decomposition rate.
  Convenient. No need to standardize the H2O2 concentration. Applicable to RBC lysates and tissue homogenates.

• Proprietary formulation increases the stability of H2O2 and catalase for more accurate results.

The Catalase Activity Assay Kit method is essentially that described by Beers & Sizer (1952) in which the decomposition of peroxide is followed spectrophotometrically at 240 nm, with modifications to increase robustness and convenience. The reaction scheme is shown below. Instead of having to calibrate precise H2O2 concentration to 10.3 mM in a tedious process, the catalase activity assay uses a certified catalase standard with known activity unit. Because catalase concentration in sample is obtained by comparing to catalase standards, calibration of precise H2O2 concentration is not necessary in this assay. Similarly, experiments can be carried out under conditions that are more convenient and more accurate. Modifications are made within the formulations to overcome problems associated with instability of diluted hydrogen peroxide and diluted enzyme standards at room temperature. There is no need to keep the reagents on ice - wasting time to bring them to the assay temperature before each individual assay.

Glutathione Assay Kit 

CAY-703002-KI01

ApoGSH Glutathione Detection Kit 

BV-K251-100

BioVisions ApoGSH Glutathione Detection Kit provides a simple in vitro assay for detecting the GSH changes in apoptosis. The assay utilizes monochlorobimane (MCB), a dye that appears to form an adduct exclusively with GSH. The unbound MCB is almost nonfluorescent, whereas the dye fluoresces blue (Ex. = 380 nm; Em. = 461 nm) when bound to glutathione. The reaction is catalyzed by glutathione S-transferase. Thus, the changes in gluotathione level in apoptosis can be easily detected using a fluorometer or a 96-well fluorometric plate reader.

ApoGSH Colorimetric Glutathione Detection Kit 

BV-K261-100

Glutathione (GSH) is the major intracellular low-molecular-weight thiol that plays a critical role in the cellular defense against oxidative stress in mammalian cells. BioVisions ApoGSH Glutathione Colorimetric Assay Kit provides a convenient, colorimetric method for analyzing either total glutathione or the reduced form of glutathione only using a microtiter plate reader. The assay is based on the glutathione recycling system by DTNB and glutathione reductase. DTNB and glutathione (GSH) react to generate 2-nitro-5-thiobenzoic acid and GSSG. Since 2-nitro-5-thiobenzoic acid is a yellow colored product, GSH concentration can be determined by measuring absorbance at 412 nm. GSH can be regenerated from GSSG by glutathione reductase, and reacts with DTNB again to produce more 2-nitro-5-thiobenzoic acid. Therefore, the recycling system dramatically improves the sensitivity of total glutathione detection. The kit includes the 5-Sulfosalicylic acid (SSA) for the removal of proteins from samples and for the protection of GSH oxidation and γ-glutamyl transpeptidase reaction. The kit can quantify glutathione from 1-100ng/well in a 200 μl reaction. For detecting lower glutathione concentrations, such as in blood samples, increasing reaction time will generate stronger signal. The kit can also specifically detect the reduced form of glutathione (GSH) by omitting the glutathione reductase from the reaction mixture. The sensitivity for detecting the reduced form of glutathione (without recycling system) is 100 times lower than detecting the total glutathione.

Glutathione Peroxidase Assay Kit 

CAY-703102-KI01

Glutathione Reductase Assay Kit 

CAY-703202-KI01

Hydrogen Peroxide (urinary) Assay Kit 

CAY-706011-KI01

oxLDL-β2GPI (human) ELISA Kit 

CAY-10007893-KI01

Lipid Hydroperoxide (LPO) Assay Kit 

CAY-705002-T100

Lipid Hydroperoxide [LPO] Assay Kit 

CAY-705003-KI01

Nitroso-thiol (RSNOs) Detection Kit 

ALX-850-037-KI01

The kit provides a complete solution for the assay of high or low MW RSNOs. The assay is divided essentially into two steps. In the first step, the sample is prepared according to which subgroup of RSNO is desired. The S-N bond is then cleaved, resulting in the formation of an equivalent of nitrite, which is assayed using the Griess reaction. The intensity of the color formed is linearly related to the concentration of nitrite, and hence, to the original RSNO concentration.

Protein Carbonyl ELISA Kit (ZenTech) 

ALX-850-312-KI01

For quantitative detection of carbonylated protein levels in plasma, other body fluids, cell and tissue extracts.

Antibody-based detection set. Advantages over colorimetric assays: Higher sensitivity - lower background - less labour-intensive - handles more samples per day.

Protein Carbonyl Assay Kit 

CAY-10005020-KI01

Superoxide Dismutase [SOD] Activity Assay Kit
ABL-SOD-560-KI01 1 Kit

• Colorimetric Assay Kit.

• Based on inhibition of hematoxylin oxidation.

• More convenient assay than the coupled cytochrome c reduction method.

• Determines total SOD activity.

• Can be used to distinguish CuZnSOD or MnSOD activity separately.

The method used in the SOD Activity Assay Kit, is essentially that described by J. P. Martin Jr., et al., with modifications to increase robustness and reliability. Briefly, autoxidation of hematoxylin (HTH2) to hematein (HT) is inhibited by SOD at the assay pH, the percentage of inhibition is linearly proportional to the amount of SOD present within a specific range. The amount of SOD in a sample is determined in the standard cytochrome c SOD unit, by measuring autoxidation rates in the presence and absence of the sample. Additional advantage of this assay is that autoxidation rate is not affected by cyanide and other reagents used to distinguish CuSOD and MnSOD activity, total SOD activity can be determined, or Cu-SOD and MnSOD activities can be determined separately by using the same kit. 

Superoxide Dismutase [SOD] Assay Kit

CAY-706002

Significant amounts of superoxide dismutase (SOD) in cellular and extracellular environments are crucial for the prevention of diseases linked to  oxidative stress. Mutations in SOD account for approximately 20% of familial  amyotrophic lateral sclerosis (ALS) cases. SOD also appears to be important in  the prevention of other neurodegenerative disorders such as Alzheimer's,  Parkinson's, and Huntington's Diseases. The reaction catalyzed by SOD is  extremely fast, having a turnover of 2 x 10^6 M-1sec-1 and the presence of  sufficient amounts of the enzyme in cells and tissues typically keeps the  concentration of superoxide very low. Quantification of SOD activity is  therefore essential in order to fully characterize the antioxidant capabilities  of a biological system. The Cayman Chemical SOD Assay kit is a fast and reliable  assay for the measurement of SOD activity from plasma, serum, tissue  homogenates, and cell lysates. SOD activity is assessed by measuring the  dismutation of superoxide radicals generated by xanthine oxidase and  hypoxanthine in a convenient 96 well format. A key feature of the kit is the  inclusion of a quality-controlled SOD standard. The standard curve generated  using this enzyme provides a means to accurately quantify the activity of all  three types of SOD (Cu/Zn-, Mn-, and Fe-SOD). Each kit contains sufficient  reagents to assay 41 samples in duplicate and includes assay buffer, sample  buffer, radical detector, SOD (Standard), xanthine oxidase, a 96 well plate, and  complete instructions.

Superoxide Dismutase [SOD]  Assay Kit 

BV-K335-100

Superoxide dismutase (SOD) is one of the most important antioxidative enzymes. It catalyzes the dismutation of the superoxide anion into hydrogen peroxide and molecular oxygen. The sensitive SOD assay kit utilizes WST-1 that produces a water-soluble formazan dye upon reduction with superoxide anion. The rate of the reduction with a superoxide anion are linearly related to the xanthine oxidase (XO) activity, and is inhibited by SOD. Therefore, the inhibition activity of SOD can be determined by a colorimetric method.

Cu/ZN-Superoxide Dismutase [SOD] (human) ELISA Kit 

BMS222

The Cu/ZnSOD ELISA is an enzyme-linked immunosorbent assay for quantitative detection of human copper zinc superoxide dismutase in cell culture supernatants, human serum, plasma, urine, amniotic fluid, fetal umbilical vein blood, or other body fluids.

Cu/ZN-Superoxide Dismutase [SOD] (human) Module Set 

BMS222MST

OXI-TEK TBARS Assay Kit 

ALX-850-287-KI01

The sensitivity of measuring thiobarbituric acid reactive substances (TBARS) has made this assay the method of choice for screening and monitoring lipid peroxidation, which is a major indication of oxidative stress. Malondialdehyde (MDA) forms a 1:2 adduct with thiobarbituric acid; the adduct can be measured by fluorometry. Biological specimens contain a mixture of TBARS, including lipid hydroperoxides and aldehydes. In practice, TBARS are expressed in equivalent numbers to MDA.

TBARS Assay Kit 

CAY-10009055-KI01

Thioredoxin Reductase Assay Kit 

CAY-10007892-KI01

NEW Stable Form of HNE (ALEXIS Biochemicals)

Yields ~ 5.2mg aldehyde after hydrolysis
For in situ production of active HNE
Allows quantitive and reproducible experiments

APF [Aminophenyl fluorescein]

Fluorescent reagent (Ex(max): 490nm; Em(max): 515nm) for the detection of highly reactive oxygen species (hROS). Immediately reacts with hROS such as hydroxyl radical, peroxynitrite and hypochlorite, and the fluorescence intensity greatly increases. Use of APF together with HPF (Prod. No. ALX-620-074) also allows for specific detection of hypochlorite (-OCl) to elucidate reliable the roles of -OCl in biological systems such as neutrophils. In addition, peroxynitrite can be detected in distinction from nitric oxide and superoxide since APF does not react with nitric oxide, superoxide and hydrogen peroxide. Moreover, APF is resistant to light-induced autooxidation. Not for sale in Japan.

ALX-620-075

2',7'-Dichlorodihydrofluorescein diacetate 

[DCDHF diacetate],  [2',7'-Dichlorofluorescein diacetate]

Cell permeable, sensitive indicator of peroxynitrite formation. After hydrolysis of the diacetate groups by cytosolic esterases or base-catalyzed cleavage of the diacetate groups, DCDHF is oxidized by peroxynitrite to the highly fluorescent product dichlorofluorescein (DCF). Formation of DCF can be monitored by fluorescence spectroscopy (Ex(max): 502nm, Em(max): 523nm), or by absorbance spectroscopy at 500nm (ε=79,500M^-1cm^-1). Neither nitric oxide, superoxide nor hydrogen peroxide alone appear to oxidize DCDHF.

ALX-610-022

Dihydrorhodamine 123 [DHR]

Sensitive indicator of peroxynitrite formation. DHR is oxidized by peroxynitrite to the highly fluorescent product rhodamine. Formation of rhodamine can be monitored by fluorescence spectroscopy (Ex(max): 500nm, Em(max): 536nm), or by absorbance spectroscopy at 500nm (ε=78'800M^-1cm^-1). Neither nitric oxide, superoxide, nor hydrogen peroxide alone appear to oxidize DHR.

ALX-610-021

HPF [Hydroxyphenyl Fluorescein]

Cell permeable fluorescent reagent (Ex(max): 490nm; Em(max): 515nm) for the detection of highly reactive oxygen species (hROS). Immediately reacts with hROS such as hydroxyl radical and peroxynitrite, and the fluorescence intensity greatly increases. In addition, peroxynitrite can be detected in distinction from nitric oxide and superoxide since HPF does not react with nitric oxide, superoxide and hydrogen peroxide. Moreover, HPF is resistant to light-induced autooxidation. HPF does not react with hypochlorite (^-OCl) either and thus can be used in combination with APF (Prod. No. ALX-620-075) which detects ^-OCl to elucidate reliably the roles of ^-OCl in biological systems such as neutrophils. Moreover, HPF is resistant to light-induced autooxidation. Not for sale in Japan.

ALX-620-074

HPF [Hydroxyphenyl Fluorescein]

CAY-10159

 
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