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Catalase Activity Colorimetric/Fluorometric Assay Kit

based on 9 citations in multiple journalsCatalase Activity Colorimetric/Fluorometric Assay Kit94.2 4
Highly Sensitive Assay, HTS
Catalog #: K773

In stock

$295.00

Product Details

Cat # +Size K773-100
Size 100 assays
Detection Method Absorbance (570 nm) or Fluorescence (Ex/Em 535/587 nm)
Species Reactivity Mammalian
Applications The kit detects high pico-unit of catalase in samples.
Features & Benefits • Simple procedure; takes ~ less than 40 minutes
• Fast and convenient
Kit Components • Catalase Assay Buffer
• OxiRed™ Probe (in DMSO)
• HRP (lyophilized)
• H₂O₂ (3%; 0.88M)
• Stop Solution
• Catalase Positive Control (lyophilized)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Catalase (EC 1.11.1.6) is a ubiquitous antioxidant enzyme that is present in nearly all living organisms. It functions to catalyze the decomposition of hydrogen peroxide (H₂O₂) to water and oxygen. BioVision’s Catalase Assay Kit provides a highly sensitive, simple, direct and HTS-ready assay for measuring Catalase activity in biological samples. In the assay, catalase first reacts with H₂O₂ to produce water and oxygen, the unconverted H₂O₂ reacts with OxiRed™ probe to produce a product, which can be measured at 570 nm (Colorimetric method) or at Ex/Em=535/587nm (fluorometric method). Catalase activity is reversely proportional to the signal. The kit detects high pico-unit of catalase in samples.


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What is the sensitivity of this assay and how can plasma/whole blood samples be processed for this assay?
The lower end of sensitivity is high pico-units to 1 miro-unit of Catalase activity. The datasheet contains instructions for Erythrocytes. Whole blood can be processed similarly. Plasma can be diluted over a range and then the dilution that gives readings within the linear range of the standard curve can be used for the assay.
Which protein assay is compatible with this kit?
We suggest you use a detergent compatible BCA assay kit: http://www.biovision.com/bca-protein-assay-kit-ii-6783.html or http://www.biovision.com/extra-sense-bca-protein-assay-kit-6785.html
What is the activity level of the positive control? How can we increase its value to be comparable with our samples?
The positive control is only a benchmark sample. As long as the values are within the range of the std curve this is fine. The positive control is not be used to compare values with the samples. The positive control is provided to validate that the assay components are all working. The customer can add more volume to get higher values but this is not necessary as long as the values are within the std. curve range. Catalase is a very vulnerable enzyme to freeze-thaw and can lose activity with storage over time.
Is sonication enough to lyse cells?
We recommend homogenizing cells. We use a glass dounce homogenizer: http://www.biovision.com/dounce-tissue-homogenizer-3034.html. This method is gentle yet effective. If needed sonication can be used in addition to homogenization. Generation of heat during sonication affects the activity of the Catalase enzyme.
Can samples from bacteria work with this kit?
Our kits are developed with mammalian samples but they work with a variety of samples from different species including bacteria. For gram positive bacteria, lysis reagents (Lysozyme treatment) might be required to rupture the cell wall. Gram negative bacterial cells can simply be homogenized as described in the protocol. We recommend testing different volumes/dilutions of the samples to make sure the final readings are within the linear range of the std curve.
Can food samples be used?
Food samples can be homogenized with the assay buffer and then centrifuged to collect the supernatant which will be the sample for the assay. Liquid food samples can be tested without any preparation. All samples must be spun down to make sure there is no floating debris or particulate material. We suggest testing several volumes of the samples to optimize the amount needed to get numbers in the linear range of the std. curve.
Can this assay be run in cuvettes?
All our assays are optimized based on 96-well plate format. If you wish to run the assays in cuvettes, less number of samples can be assayed since the volume of the cuvette will need scaling up all components.
The RFU values are same for increasing volumes of our sample. Why?
The classic cue to saturation is that when you add more sample the values decrease, meaning the maximum has already been attained and there is limitation of either reagents or Vmax has been reached already. When there is very high amount of catalase in the sample, all the substrate is quickly converted into product and then substrate is no longer available limiting the color development. When you dilute the sample, there is less catalase and hence the substrate is gradually converted to product showing a gradual increase over time. Sample volume needs to be optimized to make sure that just enough is used to get values int he linear range of the std. curve, not too high or too low.
Can individual components of this kit be purchased separately?
Yes, any of the kits components can be purchased separately without having to buy the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can alternate buffers be used for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
Is it essential to make a standard curve for every expt, or is one curve/kit enough?
Yes, it is strongly recommended to do the standards every time you do the expt. There is always a chance that something was done differently that day and we do not want any conditions to differ between standards and samples.
Héritier, L. et al., (2017) Oxidative stress induced by glyphosate-based herbicide on freshwater turtles, Environmental Toxicology and Chemistry, Aug.2017,
Baysal, M et al., (2017) Reproductive toxicity after levetiracetam administration in male rats: Evidence for role of hormonal status and oxidative stress, PLoS One, 2017, 12(4): e0175990
Liu, Tao et al. (2017) Lutein protects against β-amyloid peptide-induced oxidative stress in cerebrovascular endothelial cells through modulation of Nrf-2 and NF-κb, Cell Biol Toxicol. 2017 Feb;33(1):57-67.
Mollica, Adriano et al. (2017) An assessment of the nutraceutical potential of Juglans regia L. leaf powder in diabetic rats, Food Chem Toxicol. 2017 Mar 30.
Parrillo, Lucia et al. (2017) Olive mill wastewater-enriched diet positively affects growth, oxidative and immune status and intestinal microbiota in the crayfish, Astacus leptodactylus, Aquaculture. 2017 Apr;473: 161-168
For more citations of this product click here
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