Log In Sign up
The largest database of trusted experimental protocols

Amplex ultrared

Manufactured by Merck Group
Visit Supplier

Amplex UltraRed is a fluorogenic substrate used for the detection and quantification of hydrogen peroxide. It is a sensitive reagent that can be used in various bioanalytical applications.

Automatically generated - may contain errors

Lab products found in correlation

Amplex ultrared, by Thermo Fisher Scientific (2 mentions) Horseradish peroxidase hrp, by Merck Group (2 mentions) Free glycerol reagent, by Merck Group (1 mentions) Isoproterenol, by Merck Group (1 mentions) Superoxide dismutase sod, by Merck Group (1 mentions) Datlab 7, by Oroboros (1 mentions) Antimycin a, by Merck Group (1 mentions) Adenosine diphosphate (adp), by Merck Group (1 mentions) Rotenone, by Merck Group (1 mentions) Carbonyl cyanide m chlorophenylhydrazone cccp, by Merck Group (1 mentions) Amplex red, by Merck Group (1 mentions) Envision multilabel plate reader, by PerkinElmer (1 mentions)

Spelling variants of this product

Amplex UltraRed Reagent (3 citations)

4 protocols using amplex ultrared

1

Glycerol Release from Adipose Tissue

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
100 mg of epididymal white adipose tissue samples were removed. Samples were cut into 20 mg sections to evaluate the glycerol diffusion from tissue to the medium better; 100 mg of tissue were incubated at 37 °C in 500 μL of Krebs Ringer buffer (KRB; 12 mM HEPES, 121 mM NaCl, 4.9 mM KCl, 1.2 mM MgSO4, 0.33 mM CaCl2) containing 2% FA-free bovine serum albumin (BSA) and 0.1% glucose in the presence or absence of 10 μM isoproterenol (Sigma). Tissue was incubated for 1 h at 37 °C in a shaking bath and then gassed with 95% O2-5% CO2. At the end of the incubation period, an aliquot of the medium was used for the analysis of glycerol. A commercially available absorbance-based enzyme assay for glycerol (Free Glycerol Reagent; Sigma) was converted to fluorescence-based detection by the inclusion of the hydrogen peroxide-sensitive dye Amplex UltraRed, as reported by Clark et al. [28 (link)].
Lombardi A., Busiello R.A., De Matteis R., Lionetti L., Savarese S., Moreno M., Gentile A., Silvestri E., Senese R., de Lange P., Cioffi F., Lanni A, & Goglia F. (2019). Absence of Uncoupling Protein-3 at Thermoneutrality Impacts Lipid Handling and Energy Homeostasis in Mice. Cells, 8(8), 916.
+ Open protocol
+ Expand
2

In Vivo Penile Oxidative Stress Measurement

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
In vivo hydrogen peroxide (H2O2) and superoxide were measured in the penis via microdialysis on anesthetized rats [32 (link),33 (link)]. The penis was freed of skin and fascia, and a linear microdialysis probe (Bioanalytical Systems, Inc., West Lafayette, IN) with a 20 kDa maximal pore size was inserted into the shaft of the penis. Microdialysis probes were perfused with saline containing 100 μM Amplex Ultrared (Molecular Probes, Eugene, OR) and 1.0 U/ml horseradish peroxidase (HRP; Sigma Aldrich, St. Louis, MO) at 1.0 μl/min with a microdialysis pump (Harvard Apparatus, Holliston, MA). Amplex Ultrared is a fluorogenic substrate, which in the presence of HRP reacts with H2O2 to produce the highly fluorescent resorufin [34 (link),35 (link)]. Three 15-minute replicate samples were collected, and fluorescence of the dialysate was measured in a fluorometer (BioRad, Hercules, CA) at 510 nm Ex/590 nm Em. 10 U/ml superoxide dismutase (SOD; Sigma Aldrich, St. Louis, MO) was then added to the perfusate, allowing for conversion of superoxide that crosses over the microdialysis membrane into H2O2, which then stimulates the conversion of Amplex Ultrared to resorufin by HRP [36 (link)]. Three 15-minute replicate samples were collected, from which fluorescence was measured. Following measurements, penile tissue was harvested, and stored at −80°C until analysis.
Karakus S., Musicki B., La Favor J.D, & Burnett A.L. (2017). Cyclic AMP-Dependent Post-Translational Modification of Neuronal Nitric Oxide Synthase Neuroprotects Penile Erection in Rats. BJU international, 120(6), 861-872.
+ Open protocol
+ Expand
3

Oxidative Phosphorylation Respiratory Control

Check if the same lab product or an alternative is used in the 5 most similar protocols
SUIT protocol was used for the analysis of oxidative phosphorylation, to study respiratory control in a sequence of coupling and pathway control states induced by multiple titrations. After equilibration of the cells into the chamber the substrates, inhibitors, or uncoupler were added according to the following protocol: rotenone (f.c. 0.5 μM) to inhibit complex I, succinate (f.c., 10 mM) to stimulate complex II, ADP (f.c. 1mM) to measure the total respiratory capacity, Carbonyl cyanide m-chlorophenyl hydrazone CCCP (f.c. 0.05 μM) to determine the state of coupling of complex III, IV, V, antimycin A (f.c. 2.5 μM) to inhibit complex III (Sigma Aldrich). Hydrogen Peroxidase (H2O2) flux was assessed simultaneously using the H2O2-sensitive probe Amplex UltraRed, by the addition of Amplex Red (f.c. 10 μM) and Horseradish-Peroxidase (HRP) (f.c. 1U/ml) (Sigma Aldrich) following the variation of the resorufin concentration while performing the SUIT assay protocol. Additions by gas-tight syringe into the sealed Oxygraph chambers. Respirometric data analysis was carried out with DatLab 7 software provided by Oroboros.
Marrocco A., Frawley K., Pearce L.L., Peterson J., O’Brien J.P., Mullett S.J., Wendell S.G., St Croix C.M., Mischler S.E, & Ortiz L.A. (2021). Metabolic Adaptation of Macrophages as Mechanism of Defense against Crystalline Silica. The Journal of Immunology Author Choice, 207(6), 1627-1640.
+ Open protocol
+ Expand
4

Extracellular H2O2 Generation in Osteoclasts

Check if the same lab product or an alternative is used in the 5 most similar protocols
The generation of extracellular hydrogen peroxide (H2O2) by WT and GILT−/− osteoclasts was assessed using the redox-sensitive reagent Amplex UltraRed (Thermo Fisher Scientific). Nonadherent osteoclast precursor cells were plated at high density (2.5 × 105 cells per well; 96-well plate) and differentiated into osteoclasts using M-CSF (15 ng/ml) and RANKL (100 ng/ml) for 6 days, as described above. The following day, 150 μl of osteoclast supernatant (or media-only control) was transferred to a fresh 96-well plate and incubated with 10 μM Amplex UltraRed reagent and HRP (2 U/ml) (Sigma-Aldrich) in the dark for 30 min at room temperature. The absolute fluorescence intensity of the resulting oxidized product (resorufin) was measured using an EnVision Multilabel Plate Reader (PerkinElmer).
Ewanchuk B.W., Arnold C.R., Balce D.R., Premnath P., Orsetti T.L., Warren A.L., Olsen A., Krawetz R.J, & Yates R.M. (2021). A non-immunological role for γ-interferon–inducible lysosomal thiol reductase (GILT) in osteoclastic bone resorption. Science Advances, 7(17), eabd3684.
+ 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!