Seahorse xfp plates

Manufactured by Agilent Technologies

The Seahorse XFp plates are a type of microplate specifically designed for use with the Seahorse XFp Extracellular Flux Analyzer. These plates are used to measure the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) of live cells in a 96-well format, providing insights into cellular metabolism and function.

Automatically generated - may contain errors

Lab products found in correlation

Oligomycin, by Cayman Chemical (2 mentions) Fluoro carbonyl cyanide phenylhydrazone (fccp), by Cayman Chemical (2 mentions) Antimycin, by Cayman Chemical (2 mentions) Seahorse xf mini instrument, by Agilent Technologies (2 mentions) Ripa buffer with protease inhibitor, by Thermo Fisher Scientific (2 mentions) Pierce bca protein assay, by Thermo Fisher Scientific (2 mentions) Mito stress test kit, by Agilent Technologies (1 mentions) Seahorse xf base medium, by Agilent Technologies (1 mentions)

3 protocols using seahorse xfp plates

Mitochondrial Function Profiling in Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols

Primary rat neurons or human i³Neurons were plated on Seahorse XFp plates (Agilent) at a density of 40,000 cells/well, then either treated with 2DG or transduced with codon-optimized lentiviruses as described in previous sections. 48 h after 2DG treatment or 4 days after transduction, the Seahorse Extracellular Flux assay was performed using the Seahorse XF Mini instrument (Agilent) according to the manufacturer’s instructions. During the assay, the cells were treated sequentially with each of the following mitochondrial toxins: oligomycin (1.5 μg/ml), FCCP (3 μM), and antimycin (1 μM) (all Cayman Chemical). Immediately after the assay, the cells were lysed with RIPA buffer with protease inhibitor (Thermo Scientific). The lysates were analyzed for total protein content using the Pierce BCA protein assay (Thermo Scientific). Extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) values were then normalized to total protein.

Nelson A.T., Cicardi M.E., Markandaiah S.S., Han J.Y., Philp N.J., Welebob E., Haeusler A.R., Pasinelli P., Manfredi G., Kawamata H, & Trotti D. (2024). Glucose hypometabolism prompts RAN translation and exacerbates C9orf72-related ALS/FTD phenotypes. EMBO Reports, 25(5), 2479-2510.

+ Open protocol

+ Expand

Mitochondrial Function Assay in iPSC-derived mDANs

Check if the same lab product or an alternative is used in the 5 most similar protocols

Day 30–50 iPSC-derived mDANs were plated according to manufacturer’s instructions on 8-well Seahorse XFp plates (Agilent) previously coated with Poly-L-ornithine and laminin. Cells were allowed to mature for 3 d before being infected with hsyn-GFP-U6-shRNAs viruses for 3 d. Viruses were then removed, and media replaced with complete N2B27 for another 2–4 d. The day of the assay, culture media was replaced with Seahorse XF base medium (Agilent) supplemented with 1 mM sodium pyruvate, 2 mM glutamine, and 10 mM glucose (pH 7.4) for 1 h at 37°C. The Mito Stress Test Kit (Agilent) was prepared according to the manufacturer’s instructions: oligomycin (1 µM); carbonilcyanide ptriflouromethoxyphenylhydrazone (FCCP; 1 µM); rotenone/antimycin A (0.5 µM). After analysis, cells were lysed in 20 µl of RIPA buffer and protein levels were quantified by Nanodrop (A280).

Jiménez-Moreno N., Kollareddy M., Stathakos P., Moss J.J., Antón Z., Shoemark D.K., Sessions R.B., Witzgall R., Caldwell M, & Lane J.D. (2023). ATG8-dependent LMX1B-autophagy crosstalk shapes human midbrain dopaminergic neuronal resilience. The Journal of Cell Biology, 222(5), e201910133.

+ Open protocol

+ Expand

Mitochondrial Function Assay in Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols

Primary rat neurons or human i³Neurons were plated on Seahorse XFp plates (Agilent) at a density of 40,000 cells/well, then either treated with 2DG or transduced with codon-optimized lentiviruses as described in previous sections. 48 hours after 2DG treatment or 4 days after transduction, the Seahorse Extracellular Flux assay was performed using the Seahorse XF Mini instrument (Agilent) according to the manufacturer’s instructions. During the assay, the cells were treated sequentially with each of the following mitochondrial toxins: oligomycin (1.5 μg/ml), FCCP (3 μM), and antimycin (1 μM) (all Cayman Chemical). Immediately after the assay, the cells were lysed with RIPA buffer with protease inhibitor (Thermo Scientific). The lysates were analyzed for total protein content using the Pierce BCA protein assay (Thermo Scientific). Extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) values were then normalized to total protein.

Nelson A.T., Cicardi M.E., Markandaiah S.S., Han J., Philp N., Welebob E., Haeusler A.R., Pasinelli P., Manfredi G., Kawamata H, & Trotti D. (2023). Glucose Hypometabolism Prompts RAN Translation and Exacerbates C9orf72-related ALS/FTD Phenotypes. bioRxiv.

+ 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?

Revolutionizing how scientists
search and build protocols!