Vinci-Biochem Srl
Via Ponte di Bagnolo, 10
50059 Vinci (Firenze) Italy
vb@vincibiochem.it
Tel:+39 0571 568 147 -
 0571 568 135
Fax:+39 0571 568132 - P. IVA 05706610481

Bioenergetics,  Glucose Metabolism,  Mitochondrial Biology​​​ 


 
 
Cayman Glucose Metabolism
 
 
Mitochondial Function Assay Cayman
Mitochondrial Function Assays . Cayman
Mitochondrial Stress Test Kit
Primary Screening Assays
Oxygen Consumption Assays
Membrane Potential Assays
Functional Assays
ETC Activity Assays
Mitochondrial Content Assay
Citrate Synthase Activity Assay
ROS Generation, 
Mitochondrial Isolation Assay
   
Cayman Glucose Metabolism
Glucose Metabolism 
Glycolysis Cell-Based Assay Kit
Oxygen Consumption and Glycolysis Dual Assay Kit, Glucose Colorimetric Assay Kit, Glucose Uptake Cell-Based Assay Kit, Glucose-6-Phosphate Dehydrogenase Activity Assay Kit
Glucose-6-Phosphate Fluorometric Assay Kit, Pyruvate Assay Kit
L-Lactate Assay Kit, Glycogen Assay Kit, Oxygen Consumption Rate Assay Kit (MitoXpress®-Xtra HS Method), Oxygen Consumption / MitoMembrane Potential Dual Assay Kit
  
  
Assay Kits for Glucose Metabolism Research
 
The metabolism of glucose is central to mammalian life. Dynamic changes in any of the steps involved in processing glucose and its derivatives contribute to a wide range of diseases. Measuring the enzymes and metabolites is pivotal to biological and medical research. Cayman offers an array of tools to make these measurements, quickly, easily, and accurately. 
 
Glucose Uptake Cell-Based Assay Kit  
Code: 600470.  Glucose metabolism is a primary source of energy and biomaterials for the maintenance of cell homeostasis. Extra glucose is stored in the muscles and liver as glycogen which is hydrolyzed to glucose and released into the blood when needed. The rate of glucose uptake in cells is dynamic and tightly regulated by hormones and/or growth factors including insulin. Cancer cells exhibit increased glucose uptake and metabolism by aerobic glycolysis in order to support a high rate of proliferation. Chemicals that block glucose uptake by cancer cells have been shown to have anti-cancer effects. Cayman’s Glucose Uptake Cell-based Assay Kit provides a convenient tool for studying modulators of cellular glucose uptake. The kit employs 2-NBDG, a fluorescently-labeled deoxyglucose analog, as a probe for the detection of glucose taken up by cultured cells. Apigenin, a flavonoid that has been reported to be an inhibitor of glucose transport, is included as a control.  600470
   
Glucose Colorimetric Assay Kit  
Code: 10009582. Glucose, a monosaccharide (or simple sugar), is the most important carbohydrate in biology. Transported via the blood stream, it is the primary source of energy for the body’s cells. Glucose level is tightly regulated in the human body. Failure to maintain blood glucose in the normal range leads to conditions of persistently high (hyperglycemia) or low (hypoglycemia) blood sugar. Diabetes mellitus, characterized by persistent hyperglycemia, is the most prominent disease related to failure of blood sugar regulation. Cayman’s Glucose Colorimetric Assay Kit provides a simple, reproducible, and sensitive tool for assaying glucose in plasma, serum, and urine. The glucose assay uses the glucose oxidase-peroxide reaction for the determination of glucose concentrations. In this assay, glucose is oxidized to d-gluconolactone with concomitant reduction of the flavin adenine dinucleotide (FAD)-dependent enzyme glucose oxidase. The reduced form of glucose oxidase is regenerated to its oxidized form by molecular oxygen to produce hydrogen peroxide. Finally, with horseradish peroxidase as a catalyst, hydrogen peroxide reacts with 3,5-dichloro-2-hydroxybenzenesulfonic acid and 4-aminoantipyrine to generate a pink dye with an optimal absorption at 514 nm.  10009582
   
Why We Needed a Better Glycolysis Kit: Building a better assay  
   
pH-XtraTM Glycolysis Cell-Based Assay Kit Exclusive  
Code: 601230. Cayman’s pH-Xtra™ Glycolysis Cell-Based Assay Kit is designed to measure the extracellular acidification rate (ECAR), which is proportional to the rate of lactate production.1 This kit combines a glucose-based respiration buffer and inhibitors along with the phosphorescent pH probe pH-Xtra™ allowing for the accurate determination of glycolytic rate in a high-throughput manner. 601230
   
Glycolysis Cell-Based Assay Kit  
Code: 600450. Cayman’s Glycolysis Cell-Based Assay Kit provides a colorimetric method for detecting extracellular L-lactate, the end product of glycolysis, in cultured cells. In the assay, lactate dehydrogenase catalyzes the oxidation of lactate to pyruvate, in which the formed NADH reduces a tetrazolium substrate (INT) to a highly-colored formazan which absorbs strongly at 490-520 nm. The amount of formazan produced is proportional to the amount of lactate released into the culture medium and can be used as an indicator of the cellular glycolytic rate. This assay can be adapted to high-throughput screening for glycolysis regulators at the cellular level. Furthermore, the non-invasive sampling leaves the cells intact and thus allows multiplex testing for additional markers.  600450
   
Oxygen Consumption/Glycolysis Dual Assay Kit  
Code: 601060. Cayman’s Oxygen Consumption/Glycolysis Dual Assay Kit is a multi-parameter approach to measure cellular oxygen consumption and glycolysis in living cells. This assay utilizes MitoXpress® - Xtra, a phosphorescent oxygen probe, to measure oxygen consumption rate, and also quantifies extracellular lactate as a readout for glycolysis. Antimycin A, an inhibitor of the mitochondrial electron transport chain, is included as a control. The kit can be used for efficient screening of compounds that modulate mitochondrial and glycolytic function in cultured cells.  601060
   
Glucose-6-Phosphate Fluorometric Assay Kit  
Code: 700750. Glucose-6-phosphate (G6P) plays a key role in the homeostatic regulation of blood glucose levels by participating in glycolysis, glycogen synthesis, gluconeogenesis, and can be metabolized to NADPH, enabling protection against oxidative damage. Disruption of G6P activity leads to glycogen storage disease type I or von Gierke’s disease, a group of inherited metabolic diseases characterized by severe hypoglycemia, growth retardation, and hepatomegaly, due to accumulation of glycogen and fat in the liver. Cayman’s Glucose-6-Phosphate Fluorometric Assay provides a fluorescence-based method for detecting G6P in tissue homogenates and cell culture samples. In the assay, G6PDH catalyzes the oxidation of G6P to 6-phospho-D-gluconate, along with the concomitant reduction of NADP+ to NADPH. NADPH reacts with the fluorometric detector to yield a highly fluorescent product which can be analyzed with an excitation wavelength of 530-540 nm and an emission wavelength of 585-595 nm.  700750
   
Glucose-6-Phosphate Dehydrogenase Activity Assay Kit  
Code: 700300. Glucose-6-phosphate dehydrogenase (G6PDH) is a cytosolic enzyme that catalyzes the first step in the pentose phosphate pathway. This pathway includes converting glucose to ribose-5-phosphate, a precursor to RNA, DNA, ATP, CoA, NAD, and FAD. The pathway also generates NADPH. G6PDH deficiency, the most common enzyme deficiency worldwide, causes a spectrum of diseases including neonatal hyperbilirubinemia, acute hemolysis, and chronic hemolysis. G6PDH activity has been shown to be upregulated in rat and mouse models of obesity, hyperglycemia, and hyperinsulinemia. Cayman’s Glucose-6-Phosphate Dehydrogenase Assay provides a fluorescence-based method for detecting G6PDH activity in a variety of samples including erythrocyte lysates, tissue homogenates, and cell culture samples.  700300
   
L-Lactate Assay Kit  
Code: 700510. L(+)-Lactate is the major stereoisomer of lactate formed in human intermediary metabolism. The lactate to pyruvate ratio reflects the redox state of the cell and describes the balance between NAD+ and NADH, which is dependent on the interconversion of lactate and pyruvate via lactate dehydrogenase (LDH). Monitoring lactate levels is, therefore, a good way to evaluate the balance between tissue oxygen demand and utilization and is useful when studying cellular and animal physiology. Cayman’s L-Lactate Assay provides a fluorescence-based method for detecting L-lactate in biological samples such as serum, plasma, blood, urine, and saliva. It can also be utilized to determine intracellular and extracellular lactate concentrations in cell culture samples. In the assay, lactate dehydrogenase catalyzes the oxidation of lactate to pyruvate, along with the concomitant reduction of NAD+ to NADH. NADH reacts with the fluorescent substrate to yield a highly fluorescent product. The fluorescent product is analyzed with an excitation wavelength of 530-540 nm and an emission wavelength of 585-595 nm.  700510
   
Pyruvate Assay Kit  
Code: 700470. Pyruvate (pyruvic acid) is a key intermediate in cellular metabolic pathways and is derived primarily from glucose via glycolysis. Abnormal blood pyruvate levels are reported in a number of disorders including shock, liver disease, congestive heart failure, diabetes mellitus, thiamine deficiency, and metabolic disorders. Cayman’s Pyruvate Assay provides a fluorescence-based method for quantifying pyruvate in biological samples such as serum, plasma, blood, urine, and saliva. It can also be utilized to determine intracellular and extracellular pyruvate concentrations in cell culture samples.  700470
   
Glycogen Assay Kit  
Code: 700480. Glycogen is a polysaccharide that is the principal storage form of glucose in animal and human cells. Glycogen is made primarily by the liver and muscles, but it can also be made by glycogenesis within the brain and stomach. Cayman’s Glycogen Assay provides a simple, reproducible, and sensitive tool for assaying glycogen from tissue. In this assay, glycogen is hydrolyzed by amyloglucosidase to form ß-D-glucose, which is then specifically oxidized to D-glucono-d-lactone by glucose oxidase forming hydrogen peroxide in the process. Hydrogen peroxide, in the presence of horseradish peroxidase, reacts with 10-acetyl-3,7-dihydroxyphenoxazine (ADHP) in a 1:1 stoichiometry to generate the highly fluorescent product resorufin which is measure at an excitation wavelength of 530-540 nm and an emission wavelength of 585-595 nm.  700480
   
Oxygen Consumption Rate Assay Kit (MitoXpress® Xtra HS Method)  
Code: 600800. The oxygen consumption rate (OCR) of cells is an important indicator of normal cellular function. It is used as a parameter to study mitochondrial function as well as a marker of factors triggering the switch from healthy oxidative phosphorylation to aerobic glycolysis in cancer cells. Oxygen consumption is traditionally measured by a cumbersome oxygen electrode, a specialized piece of equipment that typically yields low sample throughput. The phosphorescent oxygen probe, MitoXpress® Xtra, developed by Luxcel Biosciences offers a novel method for analyzing oxygen consumption in whole cells. Cayman’s cell-based Oxygen Consumption Rate Assay Kit (MitoXpress® Xtra HS Method) utilizes this newly developed phosphorescent oxygen probe to measure oxygen consumption rate in living cells. Antimycin A, an inhibitor of the mitochondrial electron transport chain, is included to be used as a positive control. Glucose oxidase is also included in the kit to be used as a reference for oxygen depletion. The kit is easy to use and can be easily adapted to high throughput screening for compounds which modulate oxygen consumption rate.  600800
   
Oxygen Consumption/MitoMembrane Potential Dual Assay Kit  
Code: 600880. Cayman’s Oxygen Consumption/Mitochondrial Membrane Potential Dual Assay Kit is a multiplex assay designed to provide a comprehensive picture of mitochondrial performance in living cells. This assay simultaneously measures both oxygen consumption rate and mitochondrial membrane potential by utilizing MitoXpress® Xtra, a phosphorescent oxygen probe, and the cationic dye, JC-1. Antimycin A, an inhibitor of the mitochondrial electron transport chain, is included to be used as a positive control. Additionally, glucose oxidase is provided as a reference for oxygen depletion. This easy-to-use kit can be adapted for high throughput screening of compounds that modulate mitochondrial function.  600880
   
ATP Detection Assay Kit - Luminescence  
Code: 700410. Cayman's ATP Detection Assay Kit - Luminescence provides a simple and effective tool for measuring total ATP levels. This assay uses firefly luciferase to convert ATP and luciferin to oxyluciferin and light. The light emitted in this reaction is directly proportional to the concentration of ATP present. Using the ATP Detection Standard, quantitative measurement of ATP content can be achieved with a dynamic range of 12 fmol to 10 pmol of ATP. 700410
   
 
Code: 705502. Aconitase is an iron-sulfur protein containing a [Fe4S4]2+ cluster that catalyzes the stereospecific isomerization of citrate to isocitrate via cis-aconitate. Whereas exposure of aconitase to oxidants renders the enzyme inactive, loss of aconitase activity in cells or in biological samples treated with pro-oxidants has been interpreted as a measure of oxidative damage. Cayman’s Aconitase Assay Kit provides a simple, reproducible, and sensitive tool for assaying aconitase from tissue homogenates or cell lysates. In this assay, citrate is isomerized by aconitase into isocitrate, which is then converted to a-ketoglutarate in a reaction catalyzed by isocitric dehydrogenase. These reactions are monitored by measuring the increase in absorbance at 340 nm associated with the formation of NADPH. The rate of NADPH production is proportional to aconitase activity.
705502
   
Bioenergetics Currents - Cayman
Mitochondria and Cellular Homeostasis: Beyond ATP Synthesis
Evaluating Mitochondria Toxicity
MitoCheck Assays
Oxygen Consumption Rate Assays
Glycolysis
Oxidative Stress
Researcher Spotlight
Cayman Bioenergetics Currents
   
   
Mitochondrial Function Assays  
   
Electron Transport Chain Activity Assays  
 
   
Mitochondrial Membrane Potential 
Mitochondrial health in flow cytometry, fluorescence microscopy, and 96-well plate formats. 
JC-1 15003
Cationic dye used to study mitochondrial integrity flow cytometry, fluorescence microscopy, and 96-well plate formats. 
 
   
Mitochondrial Isolation 
• Isolate intact, functional mitochondria from heart, liver, kidney, or muscle tissue
• Complete package of reagents and step-by-step instructions
• Isolated coupled mitochondria are suitable for use in Oxygen consumption experiments and All Cayman MitoCheck Activity Assays
 
   
Citrate Synthase Activity 
• Simple and convenient determination of citrate synthase activity from isolated mitochondria or cell homogenates
• Measures the production of SH-CoA by monitoring the absorbance of Citrate Synthase Developing Reagent at 412 nm in a convenient 96-well format
 
   
   
 
   
 
Special Tools and References:
 
 
 
  
 
Isolated mitochondria versus cells
 
 
 
 
 
 
 
 
 
 
 
 
 
 




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    Vinci-Biochem Srl
    Via Ponte di Bagnolo, 10
    50059 Vinci (Firenze) Italy
    vb@vincibiochem.it
    Tel:+39 0571 568147
    Fax:+39 0571 568132
    P. IVA 05706610481