New Microplate-Based Method to Measure OCR and ECA Under Steady-State [O2]
Ann Arbor, MI, April 26, 2017 --(PR.com)-- Cayman Chemical, in collaboration with BMG LABTECH GmbH, has developed a novel assay kit to measure mitochondrial function under a dynamic range of oxygen concentrations. The Mito Stress Test (ACU Format) is designed to measure oxygen consumption rate (OCR) and extracellular acidification (ECA) simultaneously and in real time under tightly controlled atmospheric conditions using open-flow respirometry.
The assay was designed to work exclusively with microplate readers equipped with an Atmospheric Control Unit (ACU). BMG LABTECH’s multifunctional CLARIOstar® microplate reader with dual injection ports, time-resolved fluorescence detection, and an ACU was used to optimize the assay protocol. The ACU integrated into the CLARIOstar® reader facilitates measurement of cellular oxygenation at a controlled [O2], while phosphorescent extracellular O2 (MitoXpress® Xtra) and fluorescent extracellular pH (pH-Xtra™) probes provided in the kit enable multi-parametric detection of electron transport chain activity and glycolytic flux.
By combining these advanced instrumentation and assay technologies, this kit eliminates the often confounding hyperoxic conditions under which most commercially available, cell-based in vitro assays are conducted. “Because 'normoxia' varies between tissue and cell type, Cayman’s kit provides a more physiologically relevant model for certain cell lines or primary cells that typically would not be exposed to ambient oxygen concentrations,” said Dr. David Hoffman, the lead developer of this kit. Rather than using a typical microchamber, the kit provides an alternative platform that relies on the difference in steady-state oxygen, which allows measurements of OCR and ECA under various oxygen concentrations that are controlled by the user.
“The fact that cells of internal tissues are not exposed to the ambient O2 tension has often been neglected for the sake of practical work and ease of use in the lab,” said Carl Peters, Application Specialist with BMG LABTECH. “This collaboration provides an easy-to-set-up system whereby physiological conditions can be effortlessly reproduced in the lab.” The ACU can fully regulate both [O2] and [CO2] at a range from 0.1–20%, which provides a means to measure mitochondrial function at atmospheric concentrations that are lower than ambient conditions. Pre-defined scripts to automatically control the CLARIOstar® are included in the kit protocol, making assay performance and subsequent data analysis an easy plug-and-play and walk-away experience. Assay results are determined based on the lifetime measurements of MitoXpress® Xtra and pH-Xtra™ in response to changes in O2 and pH. One-click assay calculation templates using BMG LABTECH’s MARS software interface make the process of data analysis nearly instant. View example data (PDF) at www.caymanchem.com/cms/caymanchem/Literature/800171.pdf
Cayman Chemical plans to develop and optimize additional assays for bioenergetics research utilizing the technology and software of the CLARIOstar®.
About BMG LABTECH – The Microplate Reader Company
BMG LABTECH is a leading global developer and manufacturer of innovative, high-quality, and reliable microplate reader instrumentation. BMG LABTECH has been committed to producing microplate readers for more than twenty five years. By focusing on the needs of the scientific community, the company’s microplate readers have earned the company the reputation of being a technology leader in the field. BMG LABTECH has developed a wide range of dedicated and multi-mode microplate readers for life sciences applications and high-throughput screening. All BMG LABTECH microplate readers are conceived, developed, assembled, and tested entirely at BMG LABTECH headquarters in Germany. Since the establishment, BMG LABTECH has expanded to offer a worldwide sales and support network with offices in the USA, UK, Australia, Japan and France.
About Cayman Chemical Company
Cayman Chemical Company helps make research possible by supplying scientists worldwide with biochemical tools used to understand cancer, neurochemistry, oxidative injury, endocrinology, atherosclerosis, and other human health challenges. Our scientists are experts in the synthesis, purification, and characterization of biochemicals ranging from small drug-like heterocycles to complex biolipids, fatty acids, and many others for use as research reagents and qualified standards. We are also highly skilled in all aspects of assay and antibody development, protein expression, crystallization, and structure determination. In addition, we offer a wide range of analytical services using LC-MS/MS, HPLC, GC, and many other techniques. Cayman performs generic drug development and production in both Ann Arbor, Michigan and Neratovice, Czech Republic.
The assay was designed to work exclusively with microplate readers equipped with an Atmospheric Control Unit (ACU). BMG LABTECH’s multifunctional CLARIOstar® microplate reader with dual injection ports, time-resolved fluorescence detection, and an ACU was used to optimize the assay protocol. The ACU integrated into the CLARIOstar® reader facilitates measurement of cellular oxygenation at a controlled [O2], while phosphorescent extracellular O2 (MitoXpress® Xtra) and fluorescent extracellular pH (pH-Xtra™) probes provided in the kit enable multi-parametric detection of electron transport chain activity and glycolytic flux.
By combining these advanced instrumentation and assay technologies, this kit eliminates the often confounding hyperoxic conditions under which most commercially available, cell-based in vitro assays are conducted. “Because 'normoxia' varies between tissue and cell type, Cayman’s kit provides a more physiologically relevant model for certain cell lines or primary cells that typically would not be exposed to ambient oxygen concentrations,” said Dr. David Hoffman, the lead developer of this kit. Rather than using a typical microchamber, the kit provides an alternative platform that relies on the difference in steady-state oxygen, which allows measurements of OCR and ECA under various oxygen concentrations that are controlled by the user.
“The fact that cells of internal tissues are not exposed to the ambient O2 tension has often been neglected for the sake of practical work and ease of use in the lab,” said Carl Peters, Application Specialist with BMG LABTECH. “This collaboration provides an easy-to-set-up system whereby physiological conditions can be effortlessly reproduced in the lab.” The ACU can fully regulate both [O2] and [CO2] at a range from 0.1–20%, which provides a means to measure mitochondrial function at atmospheric concentrations that are lower than ambient conditions. Pre-defined scripts to automatically control the CLARIOstar® are included in the kit protocol, making assay performance and subsequent data analysis an easy plug-and-play and walk-away experience. Assay results are determined based on the lifetime measurements of MitoXpress® Xtra and pH-Xtra™ in response to changes in O2 and pH. One-click assay calculation templates using BMG LABTECH’s MARS software interface make the process of data analysis nearly instant. View example data (PDF) at www.caymanchem.com/cms/caymanchem/Literature/800171.pdf
Cayman Chemical plans to develop and optimize additional assays for bioenergetics research utilizing the technology and software of the CLARIOstar®.
About BMG LABTECH – The Microplate Reader Company
BMG LABTECH is a leading global developer and manufacturer of innovative, high-quality, and reliable microplate reader instrumentation. BMG LABTECH has been committed to producing microplate readers for more than twenty five years. By focusing on the needs of the scientific community, the company’s microplate readers have earned the company the reputation of being a technology leader in the field. BMG LABTECH has developed a wide range of dedicated and multi-mode microplate readers for life sciences applications and high-throughput screening. All BMG LABTECH microplate readers are conceived, developed, assembled, and tested entirely at BMG LABTECH headquarters in Germany. Since the establishment, BMG LABTECH has expanded to offer a worldwide sales and support network with offices in the USA, UK, Australia, Japan and France.
About Cayman Chemical Company
Cayman Chemical Company helps make research possible by supplying scientists worldwide with biochemical tools used to understand cancer, neurochemistry, oxidative injury, endocrinology, atherosclerosis, and other human health challenges. Our scientists are experts in the synthesis, purification, and characterization of biochemicals ranging from small drug-like heterocycles to complex biolipids, fatty acids, and many others for use as research reagents and qualified standards. We are also highly skilled in all aspects of assay and antibody development, protein expression, crystallization, and structure determination. In addition, we offer a wide range of analytical services using LC-MS/MS, HPLC, GC, and many other techniques. Cayman performs generic drug development and production in both Ann Arbor, Michigan and Neratovice, Czech Republic.
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Cayman Chemical Company
Bryanna Dadabbo
(734) 975-3913
www.caymanchem.com
Bryanna Dadabbo
(734) 975-3913
www.caymanchem.com
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