Log In Sign up
The largest database of trusted experimental protocols

Seahorse xf24 3 extracellular flux analyzer

Manufactured by Agilent Technologies
Visit Supplier
Sourced in United States

The Seahorse XF24-3 extracellular flux analyzer is a laboratory equipment designed to measure the oxygen consumption rate and extracellular acidification rate of cells in real-time. It provides precise measurements of cellular metabolic activity.

Automatically generated - may contain errors

Lab products found in correlation

Bca protein assay kit, by Beyotime (1 mentions) Xf pmp reagent, by Agilent Technologies (1 mentions) Sodium pyruvate, by Thermo Fisher Scientific (1 mentions) Seahorse xf24 sensor cartridge, by Agilent Technologies (1 mentions) Seahorse xf24 cell culture microplate, by Agilent Technologies (1 mentions) Seahorse xf calibrant solution, by Agilent Technologies (1 mentions) Xf base medium, by Agilent Technologies (1 mentions) Islet capture plate, by Agilent Technologies (1 mentions)

Spelling variants of this product

XF24 Analyzer (260 citations) Seahorse XF24 analyzer (184 citations) Seahorse XF24 (117 citations) Extracellular Flux Analyzer (86 citations) XF24 Flux Analyzer (37 citations) Seahorse XF24 Flux Analyzer (35 citations) XF24-3 Extracellular Flux Analyzer (30 citations) XF24 extracellular analyzer (28 citations) XF24-3 Analyzer (10 citations) Seahorse XF24 extracellular analyzer (2 citations)

6 protocols using seahorse xf24 3 extracellular flux analyzer

1

Mitochondrial Respiratory Profiling using Seahorse

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Oxygen consumption rate (OCR), extracellular acidification rate (ECAR) and mitochondria stress tests were measured using the Seahorse XF243 Extracellular Flux Analyzer (Seahorse Bioscience; Billerica, MA), as described previously (13 (link)). Cells were plated at a density of 40,000 cells/well and media replaced with XF media the following day 1hr prior to the assay. Three measurements of OCR and ECAR were taken at baseline and after each injection of the following mitochondrial stress test compounds: oligomycin (1µM; complex V inhibitor); FCCP (0.75µM; proton gradient uncoupler); antimycin A (1µM; complex III inhibitor). Basal and maximal respiration were normalized by subtracting non-mitochondrial OCR (i.e. after Antimycin A addition). Respiratory reserve capacity was calculated as the difference between maximal and basal OCR. ATP-linked OCR was derived as the difference between basal and Oligomycin A inhibited OCR. Data was normalized to total protein content in each well.
Hemachandra L.P., Shin D.H., Dier U., Iuliano J.N., Engelberth S.A., Uusitalo L.M., Murphy S.K, & Hempel N. (2015). Mitochondrial superoxide dismutase has a pro-tumorigenic role in ovarian clear cell carcinoma. Cancer research, 75(22), 4973-4984.
+ Open protocol
+ Expand
2

Mitochondrial Respiration Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
ECAR and OCR were measured using the Seahorse XF24-3 Extracellular Flux Analyzer per the manufacturer’s instructions (Seahorse Bioscience) in response to 1 µM oligomycin, 2 µM carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone, and 1 µM rotenone.
Karmaus P.W., Herrada A.A., Guy C., Neale G., Dhungana Y., Long L., Vogel P., Avila J., Clish C.B, & Chi H. (2017). Critical roles of mTORC1 signaling and metabolic reprogramming for M-CSF–mediated myelopoiesis. The Journal of Experimental Medicine, 214(9), 2629-2647.
+ Open protocol
+ Expand
3

Adipocyte Oxygen Consumption Measurement

Check if the same lab product or an alternative is used in the 5 most similar protocols
ADSCs derived from adipose tissue were directly seeded in the plate and induced to differentiation with the WA- or BA-induction procedure using the methods described previously. At day 14 of differentiation, adipocyte oxygen consumption was measured at 37°C using the Seahorse XF24-3 extracellular flux analyzer (Agilent, USA). Adipocytes were incubated in XF-assay medium containing 25 mM of glucose, 1 mM of sodium pyruvate and 2 mM of GlutMAX. Basal oxygen consumption rate (OCR) measurements were taken before sequential addition of reagents, 5 μM FCCP was added to render the maximal OCR. OCR values were normalized by the total cell protein concentration measured by BCA Protein Assay Kit (Beyotime, China).
Li H., Shen L., Zhang L., Yan B., Sun T., Guo F, & Yin X. (2020). Reduced Beige Adipogenic Potential in Subcutaneous Adipocytes Derived from Obese Chinese Individuals. Diabetes, Metabolic Syndrome and Obesity: Targets and Therapy, 13, 2551-2562.
+ Open protocol
+ Expand
4

Respirometry of Brown Adipocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols
Experiments were performed as previously described in detail (Mahdaviani et al, 2015). In brief, differentiated primary brown adipocytes were permeabilized using 5 nM XF PMP reagent (Agilent Technologies, Santa Clara, CA). Respirometry assay was performed in MAS buffer (660 mM mannitol, 210 mM sucrose, 30 mM KH2PO4, MgCl2, HEPES, EGTA, 1 mM GDP, 1% (w/v) fatty acid‐free BSA). The following substrates were used: 5 mM pyruvate, 0.5 mM or 3 mM malate, 5 mM succinate, 2 µM rotenone, 0.1 mM palmitoyl‐CoA, 0.5 mM carnitine, 0.1 mM palmitoyl‐carnitine, 5 mM ADP. The following compounds were injected: 5 μM oligomycin, 8 μM antimycin a, 0.5 mM N,N,N′,N′‐Tetramethyl‐p‐phenylenediamine (TMPD), 1 mM ascorbic acid, 50 mM sodium azide. Oxygen consumption rates were measured using the Seahorse XF24‐3 extracellular flux analyzer (Agilent Technologies, Santa Clara, CA).
Veliova M., Ferreira C.M., Benador I.Y., Jones A.E., Mahdaviani K., Brownstein A.J., Desousa B.R., Acín‐Pérez R., Petcherski A., Assali E.A., Stiles L., Divakaruni A.S., Prentki M., Corkey B.E., Liesa M., Oliveira M.F, & Shirihai O.S. (2020). Blocking mitochondrial pyruvate import in brown adipocytes induces energy wasting via lipid cycling. EMBO Reports, 21(12), e49634.
+ Open protocol
+ Expand
5

Mitochondrial Stress Testing of Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Oxygen consumption rate (OCR) was measured on a Seahorse XF243 extracellular flux analyzer (Agilent) according to manufacturer protocol for mitochondrial stress testing. Briefly, 1×104 cells/well were seeded on a Seahorse XF24 Cell Culture Microplate (Agilent) in primary mouse culture medium overnight; before running the assay, medium was changed to fresh pre-warmed mitochondrial stress test medium (XF Base Medium [Agilent], 1mM sodium pyruvate [Fisher], 10mM glucose [Sigma-Aldrich]) and cell were incubated at 37 °C in a non-CO2 incubator for 1h. Mitochondrial stress test reagents (Agilent) were diluted in mitochondrial stress test medium and loaded into individual ports of a Seahorse XF24 Sensor Cartridge (Agilent) that had been hydrated in Seahorse XF Calibrant Solution (Agilent) overnight at 37 °C. Final concentrations of stress test reagents were as follows: 8 μM oligomycin, 2.7 μM FCCP, 10 μM antimycin/rotenone. OCR was recorded and calculated for baseline respiration and maximal respiration, reserve respiration and proton leak. Measurement of total protein was also carried out subsequent to recording of the OCR and confirmed to be similar in all treatment conditions.
Duan L., Ramachandran A., Akakpo J.Y., Woolbright B.L., Zhang Y, & Jaeschke H. (2019). MICE DEFICIENT IN PYRUVATE DEHYDROGENASE KINASE 4 ARE PROTECTED AGAINST ACETAMINOPHEN-INDUCED HEPATOTOXICITY. Toxicology and applied pharmacology, 387, 114849.
+ Open protocol
+ Expand
6

Measuring Mitochondrial Respiration in Tissues

Check if the same lab product or an alternative is used in the 5 most similar protocols
At the end of the whole experiment, pWATs and sBAT were collected right after mice subjected to the different treatments described in the text and figure legends. Briefly, tissues (15–20 mg) were quickly rinsed in ice-cold PBS and loaded in the Islet capture plate (Agilent, Santa Clara, CA) containing Seahorse XF Assay Media DMEM (Agilent, Santa Clara, CA). The Seahorse XF24–3 Extracellular Flux Analyzer (Agilent Technologies) was used to measure mitochondrial respiration activity in animal WAT, BAT, and liver. We used the XF base medium with 10 mM glucose, 2.0 mM glutamine, and 1.0 mM pyruvate. Following an equilibration step, we recorded basal oxygen consumption rates (OCR, pMoles/min) by a 3-min mix, 2-min wait, and 3-min measure (looped 3 times) cycles before a digitonin (1 μg/ml) injection. Digitonin injection was used to increase tissue permeability. A 15-loops of OCR measurement was followed. The OCR data was normalized to protein weight.
Zhang J., Hou Y., Du X.L., Chen D., Sui G., Qi Y., Licinio J., Wong M.L, & Yang Y. (2020). ADORA1-driven brain-sympathetic neuro-adipose connections control body weight and adipose lipid metabolism. Molecular psychiatry, 26(7), 2805-2819.
+ 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!