| Field | Specification |
|---|---|
| Species | |
| Applications | |
| Sample type(s) | Serum, citrate-plasma, urine, cell lysate, culture media etc |
| Detection limit | |
| Detection method | |
| Plate format | |
| Assay time | |
| Storage | |
| Shelf life | |
| Shipping | |
| Catalog no. (Mfr.) | |
| Main SKU |
Overview
For quantitative determination of peroxide and evaluation of drug effects on peroxide metabolism. The assay uses OD585nm for signal readout. Compatible sample input includes Serum, citrate-plasma, urine, cell lysate, culture media etc. Typical stated assay timing is 30 min.
Key elements and design rationale
- Readout format: OD585nm supports plate-based signal acquisition and consistent comparison across matched samples.
- Sample compatibility: The stated sample scope includes Serum, citrate-plasma, urine, cell lysate, culture media etc, which is useful when aligning matrix type with calibration and control design.
- Analytical range context: The supplied specifications include a stated detection limit of 0.2 µM for interpreting low-signal samples.
- Feature emphasis: Sensitive and accurate. Enhanced color intensity using sorbitol. Detection range 0.2 µM (7 ng/mL) to 30 µM (1,020 ng/mL) H 2 O 2 in 96-well plate assay.
Additional feature notes highlight Simple and high-throughput. The procedure involves addition of a single detection reagent and incubation for 30 min. Can be readily automated as a high-throughput assay in 96-well plates for thousands of samples per day. Available format information for this listing includes 250 Tests.
Biological background
This product is centered on measurement of peroxide within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.
More details
Peroxide (e.g. hydrogen peroxide H2O2) is one of the key reactive oxygen species formed under oxidative stress conditions. High levels of peroxide formation have been linked to pathological conditions such as ageing, asthma, diabetes, atherosclerosis, cataract, inflammatory arthritis and neurodegenerative diseases. Simple, direct and automation-ready procedures for quantitative determination of peroxide find wide applications in research and drug discovery. BioAssay Systems peroxide assay kit is designed to measure peroxide concentration in biological samples without any pretreatment. The improved method utilizes the chromogenic Fe3+-xylenol orange reaction, in which a purple complex is formed when Fe2+provided in the reagent is oxidized to Fe3+by peroxides present in the sample. The intensity of the color, measured at 540-610nm, is an accurate measure of the peroxide level in the sample. The optimized formulation substantially reduces interference by substances in the raw samples.
Detection method
Colorimetric (OD 585 nm).
Detection limit and analytical sensitivity
Reported detection limit: 0.2 µM.
Procedures and timing
Stated procedure or timing information: 30 min.
Research relevance and current trends
- Plate-based quantification and side-by-side group comparison remain central use cases for this assay format.
- The product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.
- The description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.
Common research applications
- Quantify peroxide in serum, citrate-plasma, urine by OD585 nm readout.
- Compare treatment or phenotype groups using matched serum, citrate-plasma, urine handling.
- Monitor time-course or pre/post changes in serum, citrate-plasma, urine across study conditions.
Interpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.
Notes for experimental interpretation
- Matrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.
- Use appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.
How can I measure peroxides in cell cultures?
1. Collect cells by centrifugation 1,000 g for 10 min at 4°C. For adherent cells, use a rubber policeman to harvest cells.
Homogenize or sonicate cell pellet in 1-2 mL of cold buffer containing 0.9% sodium chloride and 5 mM potassium phosphate, pH 7.4.
3. Centrifuge at 10,000 x g for 10 min at 4°C.
4. Use the clear supernatant for assay. If not assaying on the same day, freeze the sample at -80°C and use it within one month.
We want to determine hydrogen peroxide in a sample that likely contains other ingredients such as surfactant, protein, propylene glycol, phosphonic acid, Tween 20, all at 1% or lower. Hydrogen peroxide is probably present at around 1%. Is this assay kit suitable?
Yes, you need to dilute your sample 1,000 to 10,000-fold in water, so that the hydrogen peroxide concentration falls within the detection range of the assay (between 10 and 100 μM). This dilution would eliminate potential interference by other ingredients such as Tween-20.
I have archived some supernatant samples from macrophages that were stimulated for 24hrs with various different stimulants. Am I right in assuming that this kit has no requirement for cells to be functional and that levels of peroxide may loiter in supernatant samples for long enough so that they can be readily measured at 24hrs?
The peroxide assay kit does not require functional cells and peroxide levels can be measured in culture media. We do not know how stable peroxide is in cell culture media and this will likely depend on the cell type and culture conditions. In general, peroxide in serum and urine samples can be detected for several days, even after freezing, but levels may change over time.
My Samples contain sodium pyruvate, is there any way I could use them with this assay?
Sodium Pyruvate below concentrations of 1 mM does not interfere with the assay, if your samples fall in this range you may simply run them. If the sodium pyruvate concentration is between 1 mM and 5 mM, you will need to run the assay with a standard curve made in the media or buffer that contains the sodium pyruvate. The assay will not have as great of detection sensitivity, but will still function. If the sodium pyruvate concentration is greater than 5 mM, the samples will not work. You will need to dilute the samples for them to work.
For laboratories requiring additional technical capacity, we provide scientific support services including assay execution, method guidance, product sourcing, and customization to align the assay with specific experimental objectives. If you need assistance selecting the appropriate kit configuration, adapting the workflow to your application, or identifying related research services, please click Talk to a Scientist, email support@biohippo.com, or review our Research Services; a member of our scientific team will follow up with recommendations tailored to your study.
Cold plasma seed priming modulates growth, redox homeostasis and stress response by inducing reactive species in tomato (Solanum lycopersicum)
Adhikari, B., et al (2020). Cold plasma seed priming modulates growth, redox homeostasis and stress response by inducing reactive species in tomato (Solanum lycopersicum). Free Radical Biology & Medicine, 156, 57-69. Assay: Peroxide in tomato leaves.
ROS-responsive and multifunctional anti-Alzheimer prodrugs: Tacrine-ibuprofen hybrids via a phenyl boronate linker
Liu, Z.,et al (2021). ROS-responsive and multifunctional anti-Alzheimer prodrugs: Tacrine-ibuprofen hybrids via a phenyl boronate linker. European Journal of Medicinal Chemistry, 212, 112997. Assay: Peroxide in antioxidant solution.
A Multifunctional Nanotherapy for Targeted Treatment of Colon Cancer by Simultaneously Regulating Tumor Microenvironment
Zhang, Q., Zhang, F., Li, S., Liu, R., Jin, T., Dou, Y. & Zhang, J. (2019). A Multifunctional Nanotherapy for Targeted Treatment of Colon Cancer by Simultaneously Regulating Tumor Microenvironment. THERANOSTICS, 9(13), 3732-3753. Assay: Peroxide in Z-VAD-FMK cells.
Insights Into the Mechanisms of Remote ischemic Preconditioning
Riedemann, I (2017). Insights Into the Mechanisms of Remote ischemic Preconditioning. PhD Dissertation the Kiel University. Assay: Peroxide in human endothelial cells.
Chemical genetics analysis of an aniline mustard anticancer agent reveals complex I of the electron transport chain as a target
Fedeles, B.I., et al. (2011). Chemical genetics analysis of an aniline mustard anticancer agent reveals complex I of the electron transport chain as a target. J Biol Chem 286(39):33910-20. Assay: Peroxide in human cancer cell.
The green tea polyphenol (-)-epigallocatechin-3-gallate ameliorates experimental immune-mediated glomerulonephritis
Peng, A., et al. (2011). The green tea polyphenol (-)-epigallocatechin-3-gallate ameliorates experimental immune-mediated glomerulonephritis. Kidney Int. 80(6):601-11. Assay: Peroxide in mouse kidney tissue, urine, serum.
H2O2-induced mitochondrial fragmentation in C2C12 myocytes
Fan, X., et al. (2010). H2O2-induced mitochondrial fragmentation in C2C12 myocytes. Free Radic Biol Med 49(11):1646-54. Assay: Peroxide in mouse cell culture supernatant.
N-(2-hydroxy phenyl) acetamide inhibits inflammation-related cytokines and ROS in adjuvant-induced arthritic (AIA) rats
Jawed, H., et al. (2010). N-(2-hydroxy phenyl) acetamide inhibits inflammation-related cytokines and ROS in adjuvant-induced arthritic (AIA) rats. Int Immunopharmacol 10(8):900-5. Assay: Peroxide in rat plasma/serum.
Phospholipase D-mTOR requirement for the Warburg effect in human cancer cells
Toschi, A., et al. (2010). Phospholipase D-mTOR requirement for the Warburg effect in human cancer cells. Cancer Lett 299(1):72-9. Assay: Peroxide in human cells.
Androgens induce dopaminergic neurotoxicity via caspase-3-dependent activation of protein kinase Cdelta
Cunningham, R.L., et al. (2009). Androgens induce dopaminergic neurotoxicity via caspase-3-dependent activation of protein kinase Cdelta. Endocrinology 150(12):5539-48. Assay: Peroxide in rat dopaminergic cell line n27.
Activation of the oxidative stress pathway by HIV-1 Vpr leads to induction of hypoxia-inducible factor 1alpha expression
Deshmane, S.L., et al. (2009). Activation of the oxidative stress pathway by HIV-1 Vpr leads to induction of hypoxia-inducible factor 1alpha expression. J Biol Chem 284(17):11364-73. Assay: Peroxide in human microglia cell line.
Hydrogen sulfide improves survival after cardiac arrest and cardiopulmonary resuscitation via a nitric oxide synthase 3-dependent mechanism in mice
Minamishima, S., et al. (2009). Hydrogen sulfide improves survival after cardiac arrest and cardiopulmonary resuscitation via a nitric oxide synthase 3-dependent mechanism in mice. Circulation 120(10):888-96. Assay: Peroxide in mouse serum.
Nguyen GN et al (2009) Drought-Induced Oxidative Conditions in Rice Anthers Leading to a Programmed Cell Death and Polle
Nguyen GN et al (2009) Drought-Induced Oxidative Conditions in Rice Anthers Leading to a Programmed Cell Death and Pollen Abortion. Journal of Agronomy and Crop Science 195(3):157-164. Assay: Peroxide in plant rice anther.
Gangliosides protect bowel in an infant model of necrotizing enterocolitis by suppressing proinflammatory signals
Schnabl, K.L., et al. (2009). Gangliosides protect bowel in an infant model of necrotizing enterocolitis by suppressing proinflammatory signals. J Pediatr Gastroenterol Nutr 49(4):382-92. Assay: Peroxide in human intestinal tissue.
Dicer ablation in oligodendrocytes provokes neuronal impairment in mice
Shin, D., et al. (2009). Dicer ablation in oligodendrocytes provokes neuronal impairment in mice. Ann Neurol 66(6):843-57. Assay: Peroxide in mouse brain tissue.
Resveratrol improves endothelial function: role of TNF{alpha} and vascular oxidative stress
Zhang, H., et al. (2009). Resveratrol improves endothelial function: role of TNF{alpha} and vascular oxidative stress. Arterioscler Thromb Vasc Biol 29(8):1164-71. Assay: Peroxide in mouse serum.
Antioxidative capacity of C(60) (buckminsterfullerene) and newly synthesized fulleropyrrolidine derivatives encapsulated in liposomes
Lens, M., et al. (2008). Antioxidative capacity of C(60) (buckminsterfullerene) and newly synthesized fulleropyrrolidine derivatives encapsulated in liposomes. Biotechnol Appl Biochem 51(Pt 3):135-40. Assay: Peroxide in liposomes.
Suppressive oligodeoxynucleotides inhibit silica-induced pulmonary inflammation
Sato, T., et al. (2008). Suppressive oligodeoxynucleotides inhibit silica-induced pulmonary inflammation. J Immunol 180(11):7648-54. Assay: Peroxide in mouse macrophages and culture supernatant.
A novel caffeine dehydrogenase in Pseudomonas sp
Yu, C.L., et al. (2008). A novel caffeine dehydrogenase in Pseudomonas sp. strain CBB1 oxidizes caffeine to trimethyluric acid. J Bacteriol 190(2):772-6. Assay: Peroxide in Pseudomonas sp. purified enzyme.
In vitro and in vivo induction of apoptosis by capsaicin in pancreatic cancer cells is mediated through ROS generation and mitochondrial death pathway
Zhang, R., et al. (2008). In vitro and in vivo induction of apoptosis by capsaicin in pancreatic cancer cells is mediated through ROS generation and mitochondrial death pathway. Apoptosis 13(12):1465-78. Assay: Peroxide in human pancreatic cancer cells.
Lactate sensitive transcription factor network in L6 cells: activation of MCT1 and mitochondrial biogenesis
Hashimoto, T., et al. (2007). Lactate sensitive transcription factor network in L6 cells: activation of MCT1 and mitochondrial biogenesis. FASEB J 21(10):2602-12. Assay: Peroxide in rat L6 cell lysate.
