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Electron Transport

Electron Transport is the process by which electrons are transported through a series of biochemical reactions, often involving the movement of electrons through a chain of carrier molecules such as proteins and enzymes.
This fundamental biological process is essential for energy production in living organisms, powering a wide range of cellular functions.
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Most cited protocols related to «Electron Transport»

1
Mitochondrial Respiration Assay
Two types of experiments are presented with isolated mitochondria. In the first, respiration by the mitochondria (5 µg/well) was sequentially measured in a coupled state with substrate present (basal respiration), followed by State 3 (phosphorylating respiration, in the presence of ADP and substrate), State 4 (non-phosphorylating or resting respiration) following conversion of ADP to ATP, State 4o (induced with the addition of oligomycin), and then maximal uncoupler-stimulated respiration (State 3u). This allows respiratory control ratios (RCR; State 3/State 4o, or State 3u/State 4o) to be assessed [18] –[20] . Unless otherwise noted, the substrate was 10 mM succinate plus 2 µM rotenone. Injections were as follows: port A, 50 µl of 40 mM ADP (4 mM final); port B, 55 µl of 25 µg/ml oligomycin (2.5 µg/ml final); port C, 60 µl of 40 µM FCCP (4 µM final); and port D, 65 µl of 40 µM antimycin A (4 µM final). The second type of experiment examined sequential electron flow through different complexes of the electron transport chain. With the initial presence of 5 µg mitochondria per well, 10 mM pyruvate, 2 mM malate and 4 µM FCCP, injections were made as follows: port A, 50 µl of 20 µM rotenone (2 µM final); port B, 55 µl of 100 mM succinate (10 mM final); port C, 60 µl of 40 µM antimycin A (4 µM final); port D, 65 µl of 100 mM ascorbate plus 1 mM N,N,N′,N′-Tetramethyl-p-phenylenediamine (TMPD, 10 mM and 100 µM final, respectively). Typical mix and measurement cycle times for the assays are illustrated in Table 1 and are common to all experiments presented unless otherwise noted.
Rogers G.W., Brand M.D., Petrosyan S., Ashok D., Elorza A.A., Ferrick D.A, & Murphy A.N. (2011). High Throughput Microplate Respiratory Measurements Using Minimal Quantities Of Isolated Mitochondria. PLoS ONE, 6(7), e21746.
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Publication 2011
Antimycin A Biological Assay Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone Cell Respiration Electron Transport malate Mitochondria Oligomycins Pyruvate Respiratory Rate Rotenone Succinate tetramethyl-p-phenylenediamine
2
Measuring Cellular Bioenergetics with Seahorse
Cellular bioenergetics of the isolated cells was determined using the extracellular flux analyzer (Seahorse Bioscience), which measures O2 and protons. This system allows for real-time, noninvasive measurements of O2 consumption rate (OCR) and proton production rate (PPR), which can be correlated to mitochondrial function/oxidative burst and glycolysis, respectively.43 (link) The injection ports attached to the wells allow for injection of inhibitors of mitochondrial respiratory chain or activators of the oxidative burst to determine the defects in individual cellular respiration pathways or enzymes. Pilot experiments for monocytes, neutrophils, platelets, and lymphocytes isolated from individual donors were performed to determine the optimal cell number required for accurate measurements of OCR and PPR. The optimum concentration of the inhibitors and activators to be used for the assessment of mitochondrial function and oxidative burst were determined by titrating the individual compounds in separate experiments against the cell number determined in the first set of experiments. First, the mean basal respiration is determined by taking 3–4 OCR measurements before the addition of the inhibitors or activators. ATP-linked OCR and proton leak were determined by injecting oligomycin at 0.5 μM (for monocytes, lymphocytes, and neutrophils) or 0.75 μM (for platelets). The fall in OCR following oligomycin injection is the rate of oxygen consumption that corresponds to ATP synthesis, and the oligomycin-insensitive rate is considered as proton leak across the inner mitochondrial membrane. FCCP, an uncoupler of the electron transport chain, was used at a concentration of 0.6 μM to determine the maximal respiration rate. This rate gives the theoretical maximum oxygen consumption that can take place at cytochrome c oxidase whether limited by availability of substrate or activity of the electron transport chain. The difference between the basal rate and this FCCP-stimulated rate is the reserve capacity of the mitochondrion, which is a measure of the maximal potential respiratory capacity the cell can utilize under conditions of stress and/or increased energetic demands. Antimycin A, an inhibitor of Complex III, was used to completely inhibit mitochondrial electron transport. The OCR determined after antimycin A injection is attributable to non-mitochondrial oxygen consumption. Mitochondrial basal respiration, proton leak, and the maximal respiration were calculated after correcting for the non-mitochondrial OCR for each assay. Cells were allowed to attach to the XF24 plate for 30–60 min before measurement of mitochondrial function. Under these conditions, viability was over 90% for all cell types and remained so over the time course of the assay. At the end of the assay period, cell lysates were collected, and OCR and PPR values normalized to the protein content in each well.
Chacko B.K., Kramer P.A., Ravi S., Johnson M.S., Hardy R.W., Ballinger S.W, & Darley-Usmar V.M. (2013). Methods for defining distinct bioenergetic profiles in platelets, lymphocytes, monocytes, and neutrophils, and the oxidative burst from human blood. Laboratory Investigation; a Journal of Technical Methods and Pathology, 93(6), 690-700.
Publication 2013
Anabolism Antimycin A Bioenergetics Biological Assay Biological Transport, Active Blood Platelets Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone Cardiac Arrest Cell Respiration Cells Donors Electrons Electron Transport Electron Transport Complex III Enzymes Glycolysis inhibitors Lymphocyte Mitochondria Mitochondrial Membrane, Inner Monocytes Neutrophil Oligomycins Oxidase, Cytochrome-c Oxygen Consumption Proteins Protons Respiratory Burst Respiratory Chain Respiratory Rate Seahorses Stress Disorders, Traumatic
3
Mitochondrial H2O2 Production Assay
Fill a cuvette with the incubation buffer (item 2, Subheading 2, see Note 13), add magnetic stirring bar, turn on the stirrer, and wait until the cuvette reaches the desired temperature (25–37°C). Add 4 U/ml of horseradish peroxidase, 10 μM Amplex Red Ultra, 40 U/ml superoxide dismutase (optional, see Note 12) and the same amount of mitochondria as used in step 3.3 to build the calibration curve. Record the fluorescence for ~150 s. Add respiratory substrates (see Note 6) and record H2O2 emission.
To illustrate a typical experimental protocol, Fig. 2 presents recordings of H2O2 production by isolated mouse brain mitochondria oxidizing NAD+-dependent substrates or succinate. The H2O2 generation is triggered by the addition of a respiratory substrate (succinate, Fig. 2a or pyruvate and malate, Fig. 2b, see Note 14). With NAD+-dependent substrates, H2O2 production was stimulated by rotenone, which inhibits NADH oxidation at Complex I (Fig. 2b). With succinate, rotenone inhibited H2O2 production indicating that it was fueled by reverse electron transfer from succinate to a site in Complex I (24 (link)). With either substrate, H2O2 production was stimulated by an inhibitor of Complex III (Antimycin A) (24 (link), 37 (link)).
, & Starkov A.A. (2010). Measurement of Mitochondrial ROS Production. Methods in molecular biology (Clifton, N.J.), 648, 245-255.
Publication 2010
Antimycin A Brain Buffers Electron Transport Electron Transport Complex III Fluorescence Horseradish Peroxidase malate Mitochondria Mus NADH NADH Dehydrogenase Complex 1 Peroxide, Hydrogen Pyruvate Respiratory Rate Rotenone Succinate Superoxide Dismutase
4
Separation of Respiratory and Glycolytic Proton Fluxes
Separation of total extracellular acidification into respiratory proton production rate (PPRresp) and glycolytic proton production rate (PPRglyc) was carried out using Eq 1 as described in (5), with the same assumptions about substrate oxidation and substrate identity.
Glycolytic rate=PPRglyc=(ECARtotBP)−(OCRtot−OCRrot/myx)(maxH+/O2)(10pH−pK11+10pH−pK1)
where ECAR = extracellular acidification rate (mpH/min), tot = total, BP = buffering power (mpH/pmol H+), OCR = oxygen consumption rate (pmol O2/min), OCRrot/myx = non-mitochondrial OCR remaining after complete inhibition of mitochondrial electron transport, max H+/O2 = the maximum H+ released to the medium per O2 consumed (and CO2 generated) by respiration, (see [5 (link)]), and K1 = the combined equilibrium constant of CO2 hydration and H2CO3 dissociation to HCO3- + H+. The overall pK for CO2(aq) + H2O → HCO3- + H+ = 6.093 at 37°C ([18 ], p. 45). The spreadsheet used for these calculations in [6 (link)] incorporates Eq 1, enabling experimental data (starting pH, buffering power, maximum H+/O2, oxygen consumption rate, and total extracellular acidification rate) to be entered and proton production rate to be calculated. This spreadsheet is available for download [6 (link)].
For this calculation, we assumed that all of the CO2 produced remained in the XF24 wells [5 (link)], and that the cells used only the supplied glucose, which was completely oxidized. For complete oxidation of glucose, 1 CO2 is produced for each O2 consumed (i.e., the respiratory quotient, RQ, = 1), and a maximum of 1 H+ is generated by the hydration and dissociation of each CO2, giving a maximum H+/O2 ratio of 1. We assumed that prior to substrate addition the cells oxidized mixed endogenous substrates, primarily glycogen. Glycogen oxidation also has maximum H+/O2 of 1, and we therefore assumed an overall RQ of 1 and maximum H+/O2 ratio of 1 for pre-substrate-addition metabolism [5 (link)]. The separation of total extracellular acidification into respiratory and glycolytic proton production rates is accurate to the extent that these assumptions are correct; if, for example, substrate oxidation was incomplete and a significant fraction of the carbon was incorporated into molecules more reduced than CO2 (such as organic acids, proteins or nucleic acids), use of the maximum H+/O2 value would overestimate glycolytic rate. If pre-substrate-addition metabolism was primarily of substrates whose RQ is less than 1, such as fatty acids, using an RQ of 1 would underestimate glycolytic rate. However, these assumptions can easily be assessed for internal consistency by post-hoc measurement of lactate produced during the experiment; under the conditions used here for C2C12 myoblasts, measured lactate production agreed quantitatively with the amounts expected from calculated glycolytic rates after correction for respiratory proton production [5 (link)], suggesting that the assumptions were essentially correct.
Mookerjee S.A., Nicholls D.G, & Brand M.D. (2016). Determining Maximum Glycolytic Capacity Using Extracellular Flux Measurements. PLoS ONE, 11(3), e0152016.
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Publication 2016
Acids Bicarbonates Carbon Cell Respiration Cells Electron Transport Fatty Acids Glucose Glycogen Glycolysis Lactates Metabolism Mitochondria Myoblasts Nucleic Acids Oxygen Consumption Proteins Protons Psychological Inhibition Respiratory Rate
5
Standardized Maternal Anthropometric Measurements
A detailed manual with instructions for all adult measurement techniques, the methods for multicentre standardisation of those measures, and the procedures for the calibration and maintenance of equipment have been published elsewhere.33 (link)
34 (link)
35 All documentation, protocols, data collection forms, and electronic transfer strategies are available at www.intergrowth21.org. Briefly, the women’s height and weight were measured in duplicate with a Seca 264 stadiometer and Seca 877 scale (Seca, Germany), respectively, on study entry between 9 and 13+6 weeks’ gestation. A first trimester body mass index (BMI) was calculated and categorised as normal weight (18.50-24.99) or overweight (25.00-29.99), according to the WHO definition.36 The same standardised methods and clinical procedures were used to measure maternal weight every five weeks (plus/minus one week) until delivery, so that the possible ranges after recruitment in which weight was measured were 14-18, 19-23, 24-28, 29-33, 34-38, and 39-42 weeks’ gestation.35
Cheikh Ismail L., Bishop D.C., Pang R., Ohuma E.O., Kac G., Abrams B., Rasmussen K., Barros F.C., Hirst J.E., Lambert A., Papageorghiou A.T., Stones W., Jaffer Y.A., Altman D.G., Noble J.A., Giolito M.R., Gravett M.G., Purwar M., Kennedy S.H., Bhutta Z.A, & Villar J. (2016). Gestational weight gain standards based on women enrolled in the Fetal Growth Longitudinal Study of the INTERGROWTH-21st Project: a prospective longitudinal cohort study. The BMJ, 352, i555.
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Publication 2016
Adult Electron Transport Index, Body Mass Mothers Obstetric Delivery Pregnancy Woman

Most recents protocols related to «Electron Transport»

1
Thin-Film Electrode Characterization
Not available on PMC !

Example 12

Different thin-film electrodes were tested using the Type 1 Linear Sweep Voltammetry Test. In more detail, thin-film electrodes formed with a stainless steel 304 (SS304) conductive layer, including an electrode with an amorphous carbon layer deposited thereon in a pure Ar atmosphere, an electrode with an amorphous carbon-containing layer deposited thereon in a 20% nitrogen atmosphere, and an electrode with an amorphous carbon-containing layer deposited thereon in a 50% nitrogen atmosphere were tested. The electrodes were all produced in a roll-to-roll sputter coater.

Anodic polarization scans in PBS, with 1 mM K4[FeII(CN)6] redox mediator added, at 25 mV/s using a saturated calomel (SCE) reference electrode and each of the SS304 electrodes as the working electrode. The results are illustrated graphically in FIG. 11. A review of FIG. 11 reveals that the electron transfer kinetics between the mediator and electrode are slightly faster when the carbon layer is sputtered in a pure Ar atmosphere, compared to a N2 containing atmosphere. However, even the films sputtered in a 1:1 Ar:N2 gas mixture is still useful in a biosensor and has an increase in deposition rate of ˜164% compared to carbon sputtered in pure Ar.

US11881549B2. Physical vapor deposited electrode for electrochemical sensors (2024-01-23). EASTMAN CHEMICAL COMPANY [US]. Inventors: Dennis Lee Ashford, II [US], Daniel Patrick Puzzo [US], Spencer Erich Hochstetler [US].
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Patent 2024
Atmosphere Biosensors calomel Carbon Electric Conductivity Electron Transport Kinetics Nitrogen Oxidation-Reduction Radionuclide Imaging Stainless Steel
2
Electrode Fabrication and Electron Transfer Kinetics
Not available on PMC !

Example 13

Different thin-film electrodes were tested using the Type 1 Cyclic Voltammetry Test. In more detail, thin-film electrodes formed with a stainless steel 304 (SS304) conductive layer and capped with a carbon containing layer sputtered in an atmosphere of N2 that ranged from 0, 5, 10, 15, 20, 40, and 50% N2 by partial pressure, respectively. The electrodes were all produced in a roll-to-roll sputter coater.

Cyclic voltammograms in PBS, with 2 mM [RuIII(NH3)6]Cl3 mediator added, at 25 mV/s using a saturated calomel (SCE) reference electrode and each of the SS304 electrodes as the working electrode. The results are illustrated graphically in FIG. 12. A review of FIG. 12 reveals that the electron transfer kinetics between the mediator and electrode are not affected by the introduction of N2 into the sputtering chamber when [RuIII(NH3)6]Cl3 is used as the redox mediator. This is unexpected because the electron transfer kinetics with a K4[FeII(CN)6] redox mediator are slightly negatively affected by the introduction of N2 into the sputtering chamber during carbon deposition.

US11881549B2. Physical vapor deposited electrode for electrochemical sensors (2024-01-23). EASTMAN CHEMICAL COMPANY [US]. Inventors: Dennis Lee Ashford, II [US], Daniel Patrick Puzzo [US], Spencer Erich Hochstetler [US].
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Patent 2024
Atmosphere calomel Carbon Electric Conductivity Electron Transport Kinetics Oxidation-Reduction Partial Pressure Stainless Steel
3
INTERGROWTH-21st: Comprehensive Fetal Growth Study
INTERGROWTH-21st was a multicentre, multiethnic, population-based project, conducted between 2009 and 2014 in eight countries. The primary aim was to study growth, health, nutrition, and neurodevelopment from less than 14 weeks’ gestation to 2 years of age. Details of the study have been described elsewhere23 (link)–25 (link). In brief, all institutions providing obstetric care in eight geographically diverse regions in Brazil, China, India, Italy, Kenya, Oman, UK, and USA were chosen as study sites. From these, healthy women with a naturally conceived, singleton pregnancy who were at low risk of adverse maternal and perinatal outcomes were prospectively enroled into FGLS, one of the main components of INTERGROWTH-21st. GA was estimated from the LMP provided that: (a) the date was certain; (b) the woman had a regular 24–32 day menstrual cycle; (c) she had not been using hormonal contraception or breastfeeding in the preceding 2 months, and (d) any discrepancy between the GAs based on LMP and CRL, measured by ultrasound at 9+0 to 13+6 weeks from the LMP was ≤7 days26 (link).
Trained, dedicated research sonographers performed ultrasound scans every 5±1 weeks using identical equipment at all sites (Philips HD9 [Philips Ultrasound, Bothell, WA, USA] with curvilinear abdominal transducers C5–2, C6-3, V7-3). We used stored images of the three standard anatomical planes: (a) fetal head in the axial view at the level of the thalami, as required for measurement of the HC; (b) abdomen in an axial view at the level of measurement of the AC, and (c) femur in the longitudinal view used for measuring FL. The detailed measurement protocol, training, standardisation, and quality-control methods, including quality scoring of images, used across all study sites are described in detail elsewhere25 (link),27 (link),28 (link) and all documentation, protocols, data collection forms, and electronic transfer strategies are freely available on the INTERGROWTH-21st website.
Lee L.H., Bradburn E., Craik R., Yaqub M., Norris S.A., Ismail L.C., Ohuma E.O., Barros F.C., Lambert A., Carvalho M., Jaffer Y.A., Gravett M., Purwar M., Wu Q., Bertino E., Munim S., Min A.M., Bhutta Z., Villar J., Kennedy S.H., Noble J.A, & Papageorghiou A.T. (2023). Machine learning for accurate estimation of fetal gestational age based on ultrasound images. NPJ Digital Medicine, 6, 36.
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Publication 2023
Abdomen Care, Prenatal Electron Transport Femur Head Hormonal Contraception Menstrual Cycle Mothers Pregnancy Thalamus Transducers Ultrasonography Woman
4
Chlorophyll Fluorescence Analysis of C. bungei
The third, fourth and fifth leaves on the top of the C. bungei seedling were selected, and the chlorophyll fluorescence parameters were measured using a modulated chlorophyll fluorescence meter (MINI-Imaging-PAM, Walz, Germany) (Gao, 2006 ; Huang et al., 2018 (link)). The maximum photochemical efficiency (Fv/Fm), actual photochemical efficiency (ФPSII), potential photochemical efficiency of PSII (Fv/Fo), non-photochemical quenching (NPQ), photochemical quenching (qP) and relative electron transport rate (rETR) through PSII were calculated (Gao, 2006 ; Huang et al., 2018 (link)).
Chen W., Mou X., Meng P., Chen J., Tang X., Meng G., Xin K., Zhang Y, & Wang C. (2023). Effects of arbuscular mycorrhizal fungus inoculation on the growth and nitrogen metabolism of Catalpa bungei C.A.Mey. under different nitrogen levels. Frontiers in Plant Science, 14, 1138184.
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Publication 2023
Chlorophyll Electron Transport Fluorescence
5
Echocardiographic Assessment of Mitral Valve Disease
Data from clinical examination, 12-lead ECG and transthoracic echocardiography (TTE) performed in our institution by experienced cardiologists within 3 months prior to surgery and at 6 months FU were available in all patients. Transthoracic echocardiograms were performed within routine clinical practice using standard methods (21 (link), 22 (link)). LV and LA diameters and volumes were recorded in the long axis parasternal and apical views, and the left ventricular ejection fraction (LVEF) was estimated visually using the Simpson biplane method. The diagnosis of MVP was made as recommended (1 (link)), and the diagnosis of flail leaflet was based on failure of leaflet coaptation with rapid systolic movement of the flail segment into the LA (23 (link), 24 (link)). MR severity was assessed following an integrative approach as recommended (22 (link)). Original data were used that were unaltered from the original prospective echocardiographic data collection by means of electronic transfer. The LV outflow tract (LVOT) diameter was measured in the parasternal long axis view, and LVOTTVI was recorded as recommended (22 (link)) by pulse wave Doppler in the apical 5-chamber view. Three cardiac cycles at least in sinus rhythm and 10 in atrial fibrillation were averaged. Stroke volume (SV) was calculated as the product of LVOT area by LVOTTVI and was indexed to body surface area (BSA) and referred to as SVi. A threshold of <35 ml/m2 was considered as a priori abnormal by reference to aortic stenosis (25 (link)). Forward LVEF was calculated as the ratio of LVOT stroke volume to LV end-diastolic volume (LVEDV), and a value <50% was considered abnormal (26 (link)).
Petolat E., Theron A., Resseguier N., Fabre C., Norscini G., Badaoui R., Habib G., Collart F., Zaffran S., Porto A, & Avierinos J.F. (2023). Prognostic value of forward flow indices in primary mitral regurgitation due to mitral valve prolapse. Frontiers in Cardiovascular Medicine, 10, 1076708.
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Publication 2023
Aortic Valve Stenosis Atrial Fibrillation Body Surface Area Cardiologists Diagnosis Diastole Echocardiography Electrocardiography, 12-Lead Electron Transport Epistropheus Movement Operative Surgical Procedures Patients Physical Examination Pulse Rate Sinus, Coronary Stroke Volume Systole Ventricular Ejection Fraction

Top products related to «Electron Transport»

1
Oxygraph 2k by Oroboros
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The Oxygraph-2k is a high-performance respirometer designed for precise measurement of oxygen consumption and production in biological samples. It provides real-time monitoring of oxygen levels, making it a valuable tool for researchers in the fields of cell biology, physiology, and bioenergetics.
2
Xf24 extracellular flux analyzer by Agilent Technologies
Sourced in United States, France
The XF24 Extracellular Flux Analyzer is a lab equipment product from Agilent Technologies. It is designed to measure the oxygen consumption rate and extracellular acidification rate of cells in real-time.
3
Pam 2500 by Walz
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The PAM-2500 is a laboratory equipment product designed for analytical purposes. It serves as a versatile tool for researchers and scientists in various fields. The core function of the PAM-2500 is to perform precise measurements and analyses, though the specific intended use may vary depending on the application.
4
Xf96 extracellular flux analyzer by Agilent Technologies
Sourced in United States
The XF96 Extracellular Flux Analyzer is a laboratory instrument designed to measure the metabolic activity of cells in a high-throughput manner. The device is capable of simultaneously assessing the oxygen consumption rate and extracellular acidification rate of cells, providing insights into their respiratory and glycolytic activity.
5
Li 6400xt by LI COR
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The LI-6400XT is a portable photosynthesis system designed for measuring gas exchange in plants. It is capable of measuring net photosynthesis, transpiration, stomatal conductance, and other physiological parameters. The system consists of a control unit and a leaf chamber that encloses a portion of a plant leaf.
6
Li 6400 by LI COR
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The LI-6400 is a portable photosynthesis system designed for measuring gas exchange in plants. It is capable of measuring net carbon dioxide and water vapor exchange, as well as environmental conditions such as temperature, humidity, and light levels.
7
Rotenone by Merck Group
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Rotenone is a naturally occurring insecticide and piscicide derived from the roots of certain tropical plants. It is commonly used as a research tool in laboratory settings to study cellular processes and mitochondrial function. Rotenone acts by inhibiting the electron transport chain in mitochondria, leading to the disruption of cellular respiration and energy production.
8
Dual pam 100 by Walz
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The Dual-PAM-100 is a laboratory instrument designed for simultaneous measurement of chlorophyll a fluorescence and P700 absorbance changes. It provides reliable data on the photosynthetic performance of plants and algae.
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Antimycin a by Merck Group
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Antimycin A is a chemical compound that acts as a potent inhibitor of mitochondrial respiration. It functions by blocking the electron transport chain, specifically by interfering with the activity of the cytochrome bc1 complex. This disruption in the respiratory process leads to the inhibition of cellular respiration and energy production within cells.
10
Oligomycin by Merck Group
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Oligomycin is a laboratory product manufactured by Merck Group. It functions as an inhibitor of the mitochondrial F1F0-ATP synthase enzyme complex, which is responsible for the synthesis of adenosine triphosphate (ATP) in cells. Oligomycin is commonly used in research applications to study cellular bioenergetics and mitochondrial function.

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