Mitoxpress xtra

Manufactured by Agilent Technologies

Sourced in United States

The MitoXpress Xtra is a fluorescent probe designed to measure oxygen consumption and mitochondrial respiration in live cells. It provides a sensitive and quantitative way to monitor oxygen levels in real-time, enabling the assessment of cellular metabolic activity.

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Lab products found in correlation

Celltiter glo, by Promega (1 mentions) Bca protein assay kit, by Thermo Fisher Scientific (1 mentions) Synergy neo2, by Agilent Technologies (1 mentions)

Spelling variants of this product

MitoXpress Xtra reagent (3 citations) MitoXpress Xtra Oxygen Consumption Assay (3 citations) MitoXpress Xtra Oxygen Consumption reagent (2 citations) MitoXpress® Xtra assay (2 citations)

6 protocols using mitoxpress xtra

Oxygen Consumption Rate Measurement

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OCR was determined by Seahorse Flux Analyzer XF96 (Agilent) according to the manufacturer's instructions. In brief, 80% of confluent cells were equilibrated for 1 h at 37 °C without CO₂. OCR was then measured before and after sequential injection of indicated compounds in the corresponding figure.
OCR observation was also determined by Agilent MitoXpress Xtra (XF200 Oxygen Consumption Assay). In brief, NCI‐H82 cells were seeded at a concentration of 80 000/90uL/well in a 96‐well plate. Then, there was addition with 10 uL Mitoexpress Xtra reagent following covered with 100 uL mineral oil. Read the plate immediately at 37 °C in a fluorescence plate reader (380 nm for excitation and 650 nm for emission) for 2 h and then collect the data.

Wu C., Liu Y., Liu W., Zou T., Lu S., Zhu C., He L., Chen J., Fang L., Zou L., Wang P., Fan L., Wang H., You H., Chen J., Fang J., Jiang C, & Shi Y. (2022). NNMT‐DNMT1 Axis is Essential for Maintaining Cancer Cell Sensitivity to Oxidative Phosphorylation Inhibition. Advanced Science, 10(1), 2202642.

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Fluorescent Probe for Neuronal Respiration

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The level of cellular oxygen consumption was measured by applying a fluorescence-based probe, MitoXpress Xtra (Agilent) as per manufacturer’s protocol. In brief, MitoXpress Xtra (10 μl) was added to neurons (90 μl media) grown in 96-well plate. Immediately after treatment, all wells were sealed by 100 μl mineral oil. Antimycin (1μM), an inhibitor of the Complex III, was used as a negative control.

Park H.A., Crowe-White K.M., Ciesla L., Scott M., Bannerman S., Davis A.U., Adhikari B., Burnett G., Broman K., Ferdous K.A., Lackey K.H., Licznerski P, & Jonas E.A. (2022). Alpha-Tocotrienol Enhances Arborization of Primary Hippocampal Neurons via Upregulation of Bcl-xL. Nutrition research (New York, N.Y.), 101, 31-42.

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Fluorescent Glycoconjugate Synthesis and Characterization

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The PtPFPP dye was from Frontier Scientific
(Inochem Ltd., Lancashire, U.K.). 1-Thio-β-d-glucopyranoside
sodium salt (1Glc), and 2-thioethyl-β-d-glucopyranoside
(2Glc) were from Carbosynth Ltd. (Berkshire, U.K.). O-(2-Carboxyethyl)-O′-(2-mercaptoethyl)heptaethylene
glycol (cPEG-SH), O-(2-mercaptoethyl)-O′-methyl-hexa(ethylene
glycol) (mPEG-SH), 2-(Boc-amino)ethanethiol (Boc-CA) were from Sigma-Aldrich.
The cellular ATP assay CellTiter-Glo was from Promega (Madison, WI).
The BCA Protein Assay kit was from Thermo Fisher Scientific (Rockford,
IL). The MitoXpress-Xtra was from Agilent (Santa Clara, CA). All of
the other reagents were from Sigma-Aldrich.

Zanetti C., Gaspar R.D., Zhdanov A.V., Maguire N.M., Joyce S.A., Collins S.G., Maguire A.R, & Papkovsky D.B. (2022). Heterosubstituted Derivatives of PtPFPP for O2 Sensing and Cell Analysis: Structure–Activity Relationships. Bioconjugate Chemistry, 33(11), 2161-2169.

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Worm Oxygen Consumption Assay

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The OCR assay was performed according to (Zuo et al., 2017) using the MitoXpress Xtra oxygen consumption assay kit (Agilent, USA). Approximately 100–150 worms were recovered from each bacterial diet plate and washed three times with S-basal solution to remove excess bacteria. The worms were then transferred to wells of a 96-well plate in a final sample volume of 90 μl. Then 10μl of the oxygen probe was added to each sample. The wells of the 96 well plate were then covered with two drops of mineral oil and immediately read using a Synergy Neo 2 plate reader using Gen5 software (BioTek, Wisnooski, VT, USA) in a time-resolved fluorescence mode with 380 nm excitation and 650 nm emission filters.

Amin M.R., Mahmud S.A., Dowgielewicz J.L., Sapkota M, & Pellegrino M.W. (2020). A novel gene-diet interaction promotes organismal lifespan and host protection during infection via the mitochondrial UPR. PLoS Genetics, 16(12), e1009234.

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Measuring Cellular Oxygen Consumption

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The level of cellular oxygen consumption was measured by applying a fluorescence-based probe, MitoXpress Xtra (Agilent) as per manufacturer’s protocol. In brief, MitoXpress Xtra (10 μl) was added to cells (90 μl media) grown in 96-well plate. Immediately after treatment, all wells were sealed with 100 μl mineral oil. Antimycin (1 μM), an inhibitor of the Complex III, was used as a negative control.

Welin Henriksson E., Wahren-Herlenius M., Lundberg I., Mellquist E, & Pettersson I. (1999). Key residues revealed in a major conformational epitope of the U1-70K protein. Proceedings of the National Academy of Sciences of the United States of America, 96(25), 14487-14492.

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Mitochondrial Complex II Respiration Assay

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Freshly isolated mitochondria (60 μg) were incubated in a reaction buffer (250 mmol/L sucrose, 10 mmol/L Tris base, 1 mmol/L MgCl₂). Mitochondrial oxygen consumption was measured using Tecan Spark multi‐mode plate reader with MitoXpress Xtra fluorescent sensor reagent (Agilent Technology, Santa Clara, CA) to measure dissolved oxygen level (Ex: 380 nm/Em: 670 nm). Complex II‐driven state III respiration was stimulated by adding 10 mmol/L Succinate and 1 μmol/L Rotenone and 2.5 mmol/L ADP. Relative fluorescent change per minute was calculated using operation software.

Kitaoka Y., Tamura Y., Takahashi K., Takeda K., Takemasa T, & Hatta H. (2019). Effects of Nrf2 deficiency on mitochondrial oxidative stress in aged skeletal muscle. Physiological Reports, 7(3), e13998.

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